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The expression of CD143 (angiotensin-I-converting enzyme, ACE) in cardiovascular diseases may be an important determinant of local angiotensin and kinin concentrations. Much of the experimental and clinical evidence suggests a crucial role for Ang II in fibrogenesis and the development of atherosclerosis. Therefore, we have studied the distribution of CD143 in atherosclerotic and non-atherosclerotic segments isolated from different parts of the human vascular tree, including aorta and coronary, carotid, brachial, renal, iliac and femoral arteries, and staged according to the AHA. Two hundred and thirty native and formalin-fixed specimens of 80 patients were analysed by sensitive APAAP-technique using ten different monoclonal and polyclonal antibodies to human CD143 and several controls. In non-atherosclerotic segments or intimal thickening, CD143 was found almost restricted to the endothelial cells of adventitial arterioles and small muscular arteries. In contrast, a striking accumulation of CD143 was detected in all early and advanced atherosclerotic lesions. This de-novo occurrence of CD143 within the intimal vascular wall was caused by spindle-shaped subendothelial cells with macrophagic/histocytic features, activated macrophages and foam cells. In addition, advanced lesions of atherosclerosis showed a marked neo-expression of CD143 in newly formed intimal microvessels. Hypocellular fibrotic plaques depleted in microvessels and macrophages showed only little CD143. The de-novo occurrence of CD143 was dependent on the stage of atherosclerosis but not on its particular localisation within the vascular system. This early and obligatory CD143 expression at an unusual vascular site may contribute to unusual tissue levels of angiotensins as indicated by co-localisation of immunoreactive Ang II. Thus, it may be an important pathogenetic step in the development of atherosclerosis and an established target for pharmacological prevention.

Isolation of single cells or cell clusters from complex tissue sections has become possible by microdissection techniques. Employing laser-assisted cell picking, cell-specific mRNA analysis of a few isolated cell profiles may be performed. However, microscopic discrimination of different cell types in routinely stained tissue sections is limited, whereas immunostaining enables a more precise access to cells of interest. This approach was noted to interfere with mRNA recovery. To define optimal conditions for mRNA amplification from immunodetected cells, we systematically investigated several potential affectors. Kind of fixation, antibodies and staining reagents, incubation and total processing time, and digestion with proteinase K turned out to influence mRNA stability. We present rapid protocols for immunohistochemistry and immunofluorescence with total incubation times of approximately 25 to 40 minutes and 10 to 20 minutes, respectively, and suggest mRNA amplification without a preceding extraction step. Applying these protocols to oligocellular clusters containing approximately 20 cell profiles and nuclei each from lung and kidney tissue, the highest efficiency rates of mRNA amplification were obtained when combining short-term formalin fixation, reduction of antibody incubation time, application of immunofluorescence, and digestion with proteinase K. Thus, the successful combination of immunostaining and laser-assisted cell picking remarkably improves cell type-specific analysis of gene expression within complex tissues.

Angiotensin-converting enzyme in non-neoplastic kidney diseases. Background The angiotensin I-converting enzyme (ACE, CD143, kininase II) plays a critical role in controlling the level of vasoactive peptides such as angiotensins and kinins in the local circulations and tissue interstitium. Because recent work has documented a vessel-, organ-, and species-specific pattern of endothelial ACE expression in the vascular system, we have analyzed whether or not changes of this pattern occur in vessels, tubules, and interstitium of the human kidney that is affected by different non-neoplastic diseases. Methods Using a set of well-characterized monoclonal antibodies (mAbs), ACE was assessed on renal tissue of 135 patients by immunohistochemistry, including an additional analysis at the ultrastructural level. A semiquantitative evaluation allowed the estimation and comparison of ACE content in different renal compartments. These data were compared with several clinical findings, diagnosis, therapeutic modalities, and histological features. Results In contrast to the normal human kidney, where ACE is abundant in the brush border of the proximal tubule but is usually absent in endothelial cells of any vessel type, an endothelial neoexpression of ACE was observed in different diseases. In general, this neoexpression was associated with histological sites of interstitial fibrosis and showed some selectivity for glomerular endothelial cells in diabetes mellitus and chronic arterial hypertension. There was also a loss of epithelial ACE in the proximal tubule in certain pathological conditions, for example, in chronic fibroplastic processes, acute pyelonephritis, and different stages of acute renal failure. Conclusions Neoexpression of ACE by renal endothelial cells, as well as changes of the tubular ACE content, is a common finding in diseased human kidneys. As associated with certain tissue sites, clinical and/or morphological features, these changes may be involved in parenchymal remodeling and renal pathophysiology.

Objective: Orthotopic heart transplantation has become an accepted therapeutic concept for adult patients with endstage heart disease. In newborns and infants this procedure is still a matter of discussion because of unknown long-term results and the lack of donor organs. Methods: Since March 1988 we have performed 40 orthotopic heart transplantation in 39 infants who were from 1 to 280 days of age. Indications for transplantation included hypoplastic left-heart syndrome ( n = 28), dilative cardiomyopathy (n = 4), endocardial fibroelastosis (n = 4) and other complex structural anomalies (n = 3). The mean waiting period for transplantation was 53 days. A donor‐recipient weight ratio up to 4.0 was accepted. Profound hypothermic circulatory arrest was used for graft implantation in all those patients who required extensive aortic arch reconstruction (71%). The initial immunomodulation was based on Cyclosporine, Azathioprine and Prednisolone. Patients who underwent transplantation during the first 6 weeks of life received a chronic single-drug therapy with Cyclosporine after 1 year. Results: There were six peri-operative deaths caused by drug-resistant right-heart failure in three cases, humoral rejection (n = 1), CMV infection (n = 1) and multi organ failure (n = 1). One infant died late, due to rejection. The actuarial survival rate for the entire group is now 82%. There is a remarkable influence of increasing experience. Whereas six of 15 infants who had heart transplantation between 1988 and 1993 died early post-operatively (survival rate: 60%), only one late death occurred among 24 recipients in the period from 1994 to April 1997 (survival rate: 96%). Episodes of rejection occurred once or several times in about half of the patients in this series (48%). All surviving children are living at home in excellent condition. Conclusions: Heart transplantation during early infancy is a rational and durable therapy for heart diseases with irreversible myocardial failure or severe structural anomalies. The intermediate-term results have been encouraging in many centers, but more data must be accumulated to determine the sequelae of chronic immunosuppression. The lack of donor organs remains one of the major problems in pediatric heart transplantation. © 1998 Elsevier Science B.V. All rights reserved

Purpose: To study the morphology of corneal tunnel incisions using diamond and steel keratomes. Setting: Department of Ophthalmology, University of Giessen, Germany. Methods: Corneal tunnel incisions were performed in six human cadaver eyes using three types of diamond keratomes and a steel keratome. The incision profile and morphology were evaluated and compared using light and scanning electron microscopy. Results: The steel keratome caused more disruption of corneal stromal tissue, while the diamond keratomes produced a more regular, smoother incision. The dissecting incision resulted in a smoother surface of cut stromal tissue than the stab incision. Conclusions: The high quality of corneal tunnel incisions produced with diamond keratomes is the result of their exceptional sharpness, which may have a beneficial effect on wound healing.

Von Juni 1988 bis Dezember 1996 wurden am Kinderherzzentrum Giesen 36 Patienten, im Alter von weniger als einem Jahr, einer Herztransplantation zugefuhrt. Zugrundeliegende Diagnosen waren ein hypoplastisches Linksherzsyndrom (n = 26), Endokardfibroelastose (n = 4), Kardiomyopathie (n = 3) und komplexe Vitien (n = 3). Die mittlere Wartezeit von Transplantation betrug 52 Tage, das Verhaltnis der Korpergewichte von Spender zu Empfanger lag im Mittel bei 1,8. Sieben Patienten (19%) verstarben nach Transplantation, uberwiegend innerhalb des ersten Monats nach der Transplantation. Die kumulative Uberlebenswahrscheinlichkeit liegt im Geamtkollektiv bei 79%. Der Einflus zunehmender Erfahrung zeigt sich beim Vergleich der Patienten mit Transplantationen von 1988 bis 1993 (n ? 15) und von 1994 bis 1996 (n = 21). Wahrend im ersten Zeitraum lediglich 50% uberlebten, betrug die Uberlebensrate der spater transplantierten Kinder 92%, die 1-Jahres-Uberlebensrate lag bei 100%. Bei 20 Patienten traten insgesamt 31 Abstosungsepisoden auf, wobei 2 Sauglinge verstarben. 71% aller Abstosungen traten innerhalb des ersten Monats nach der Transplantation auf. Die Nierenfunktion ist ein Jahr nach der Transplantation nur masiggradig eingeschrankt ohne Tendenz zur Verschlechterung. Die somatische Entwicklung verlauft bei nahezu allen Kindern perzentilengerecht. Die Lebensqualitat der Patienten nach der Transplantation ist ausgezeichnet. Alle Patienten leben ohne Einschrankungen zu Hause. Bei zwei Patienten liegt allerdings ein neurologisches Defizit vor. Bislang gibt es im Gesamtkollektiv keine Anzeichen fur eine Transplantatvaskulopathie oder eine maligne Erkrankung. Eine Herztransplantation stellt nach diesen Ergebnissen eine uberdenkenswerte Alternative in der Behandlung sehr komplexer Vitien und Kardiomyopathien im Neugeborenen- und Sauglingsalter dar.

PROBLEM: The invasion of trophoblast cells into the uterine wall and its arterial system is essential for the normal development of pregnancy. Cell adhesion molecules (CAM), such as the immunoglobulin superfamily and integrins, play a crucial role in a number of immunological reactions and in the invasion of the human trophoblast. Intrauterine growth restriction (IUGR) has been associated with abnormal trophoblast invasion. Therefore, the expression of CAM in the extravillous trophoblast of pregnancies complicated by IUGR might be different from normal pregnancies. METHOD OF STUDY: Normal (n = 21) and IUGR (n = 19) placentas were collected and stored at -70 degrees C. Immunohistochemistry (avidin-biotin complex peroxidase-doublestaining) of frozen tissue sections was performed using antibodies specific for the immunoglobulin superfamily vascular adhesion molecule-1 (VCAM-1;CD 106), intercellular adhesion molecule (ICAM-1) (CD 54), ICAM-2 (CD 102), ICAM-3 (CD 50), the integrins alpha 2 beta 1, alpha 3 beta 1, alpha 4 beta 1, alpha 5 beta 1, alpha 6 beta 1 and cytokeratin. The percentage of immunopositive extravillous trophoblast cells (EVT) and the intensity of the immunoreactivity for the various CAM and integrin antibodies was assessed. RESULTS: In IUGR placentas, there was less expression of VCAM-1 (CD 106), alpha 2 beta 1, alpha 3 beta 1, and alpha 5 beta 1 (P < 0.05) in the extravillous trophoblast than in normal pregnancies. Finally we observed for the first time that ICAM-3 was expressed on EVT and that its expression was markedly up-regulated in the EVT of IUGR placentas. No differences were found for ICAM-1 (CD 54), ICAM-2 (CD 102), alpha 4 beta 1 and alpha 6 beta 1. CONCLUSION: Our data show that there are significant differences in the expression of cell adhesion molecules of the extravillous trophoblast from IUGR and normal pregnancies. These differences might reflect changes in the immunological reactions and cell-cell interactions between mother and the developing fetus which could interfere with fetal growth.

SummaryAortic atresia is rare in the setting of a normally developed left ventricle with a ventricular septal defect. In this combination, as far as we know, it has been described only with concordant ventriculo-arterial connections, for which seven one-stage biventricular repairs have now been described. We describe here the case of a 3½-month-old male infant with a similar combination but with discordant ventriculo-arterial connections and severe pulmonary hypertension. It was the right ventricle which achieved normal size in this arrangement, presumably because of the ventricular septal defect. One-stage biventricular correction was accomplished, employing a single pulmonary allograft, aided by massive doses of Prostaglandin El. As far as we know this is the first report of the combination of aortic atresia, discordant ventriculo-arterial connections, a ventricular septal defect and balanced ventricles. We complement our surgical account, nonetheless, with a description of a comparable specimen from our anatomic museum.

Studies in different preparations have suggested that Na+/H+ exchange is one mechanism causally involved in cell death in myocardial ischemia and reperfusion. The time delay of cell death by pretreatment with the Na+/H+-exchange inhibitor HOE642, cariporide (4-isopropyl-3-methylsulphonylbenzoyl-guanidine methanesulphonate), was investigated in regionally ischemic, reperfused porcine hearts. HOE642 (1 mg/kg) was injected intravenously in 14 thoracotomized pigs 10 min before occlusion of the left anterior descending coronary artery (45 min of ischemia, six pigs; 70 min of ischemia, six pigs; 90 min of ischemia, two pigs). Ischemia was followed by 24 h of reperfusion. Six animals (45 min of ischemia) served as controls. Infarct size was determined as a ratio of infarcted (tetrazolium stain, histology) to ischemic myocardium (dye technique), and regional myocardial function was assessed by sonomicrometry. HOE642 did not affect global hemodynamic parameters. In the pretreated group with 45 min of ischemia, HOE642 significantly decreased histochemical infarct size from 51.2 +/- 12.6% (control group) to 13.2 +/- 12% (p < 0.005) and histologic infarct size from 44.5 +/- 9% to 17.1 +/- 7% (p < 0.005). Recovery of regional systolic shortening after 24 h of reperfusion was improved from 2 +/- 6% (control group) to 12 +/- 7% (p = 0.02). In addition, myocardial contracture and increase in heart rate during early reperfusion were attenuated. When ischemia was prolonged to 70 min after pretreatment with HOE642, infarct size, recovery of systolic shortening, myocardial contracture, and increase in heart rate did not differ from those of the control group. Pretreatment with HOE642 increased the tolerance to ischemia/reperfusion by approximately 20-25 min. Inhibition of Na+/H+ exchange appears to be very promising in the clinical treatment of acute myocardial ischemia and reperfusion.

Besides the degree of carotid artery stenosis, the composition of the plaque may help to predict the thromboembolic risk. Low echogenicity on ultrasound and hemorrhage into the atheroma demonstrated histopathologically have been shown to be associated with a higher risk of embolism. Twenty-nine consecutive patients with carotid artery stenosis and scheduled for carotid endarterectomy were investigated preoperatively by B-mode ultrasound. Post-operatively the endarterectomy specimens were examined histopathologically. Neither atheroma with hemorrhage nor atheroma without hemorrhage were significantly associated with echolucent ultrasound presentation. Out of the 10 lesions echolucent and homogeneous on ultrasound, six corresponded to atheroma with hemorrhage, two corresponded to atheroma with hemorrhage plus thrombus, two corresponded to fibrous plaque plus thrombus, and one corresponded to pure thrombus. Out of the 4 lesions heterogeneous and predominantly echolucent, one corresponded to atheroma without hemorrhage plus thrombus, one corresponded to atheroma with hemorrhage, one corresponded to atheroma with hemorrhage plus thrombus, one corresponded to atheroma with hemorrhage plus fibrous plaque. Seven out of the 18 atheromas with hemorrhage did not present as purely or predominantly echolucent lesions, six of them were even homogeneously echogenic. Plaque surface could not reliably be predicted by ultrasound. In our study, there was no significant correlation between ultrasound and histology of the lesion.

Plasma and tissue concentrations of the angiotensin-I converting enzyme (ACE) have been shown to be associated with the ACE insertion/deletion (I/D) polymorphism. The purpose of this study was to examine the relation of ACE levels in atherosclerotic plaques to the ACE I/D polymorphism and to restenosis after balloon angioplasty and directional atherectomy (DCA). The study included 104 patients who underwent DCA and received angiographic follow-up at 12 to 18 months. The amount of ACE protein in various morphologically defined plaque components (fibrous, atheromatous, and complicated lesions) of the atherectomy specimens was determined by qualitative and semiquantitative immunohistochemistry. ACE levels were related to the ACE genotype, to plaque morphology and to the risk of restenosis. Sequential staining revealed that pathologic ACE overexpression of the atherosclerotic lesions occurred in intimal smooth muscle cells, fibrocytes/fibroblasts and macrophage/foam cells. The ACE content of the whole plaques and of the single plaque components was not associated with the I/D polymorphism, but with restenosis after coronary interventions. In addition, ACE levels in the atherosclerotic lesions correlated with the severity of vessel wall damage. The ACE phenotype might serve as an indicator for the risk of restenosis after coronary interventions.

Many of the antigens commonly investigated in histopathology can be enhanced by microwave pretreatment (MWPT) of formalin fixed, paraffin embedded tissue sections. We developed a double labeling method using microwave heating to detect otherwise undetectable nuclear antigens combined with immunohistochemistry (IHC) of cytoplasmic or membranous antigens that do not benefit from MWPT. We used the same primary antibody solutions used in single antibody IHC. The staining technique is based on the alkaline phosphatase anti-alkaline phosphatase (APAAP) and the labeled avidin-biotin (LSAB) methods. Four different protocols were tested, each modifying the sequence of MWPT, APAAP and LSAB staining. In this study Ki67, estrogen receptor, progesterone receptor, c-neu, CD68 and desmin primary antibodies were used in routinely formalin fixed, paraffin embedded tissues of 50 tumor specimens. MWPT followed by LSAB for microwave enhanced antigens and APAAP for antigens that cannot be enhanced by MWPT gave the best double staining results. This method improves characterization of tumor cell features from paraffin embedded tissue and should aid analysis of tumor differentiation, receptor status and nuclear proteins in the single cells in archival tissues.

Second primaries in the upper aerodigestive tract (UADT) are increasingly related to bad prognosis. So early detection of small cancer foci is required to improve the survival rate. Here, autofluorescence (AF) might become useful as in vivo biochemical changes of cancer metabolism can be shown by fluorescence.For more exact data on AF in cancerous lesions 32 cancer specimens of the UADT and 57 specimens of normal oral mucosa as a control group were examined by using autofluorescent microscopy and spectroscopy, the tissue being activated with light at 365 nm wavelength. Video-controlled in vivo fluorescence examinations in field cancerisation of the UADT were also investigated and compared with pathohistological findings.The intensity of AF of the connective tissue was observed to increase at the tumour border. Contrary to results in the literature, no homogeneous fluorescence gradient could be proved between marginal epithelium and the tumour itself. The brightly shining submucosal elastic fibres allowed fluorescent spectroscopic detection of the tumour margins in vitro in 71%. Even small precancerous lesions were traced in vivo mainly by keratinization when using the autofluorescence diagnostic imaging system.Our examination proved a multi-factorial genesis of autofluorescence with strong inter-individual variations. Preliminary clinical examination showed that this method can be applied as an additional tool for early detection of cancerous lesions in the UADT.

To investigate the efficacy and safety of mechanical thrombectomy for iliac vein thrombosis using Rotarex and Aspirex catheters in a pig model. Iliac vein thrombosis was induced in six pigs by means of an occlusion-balloon catheter and thrombin injection. The presence of thrombi was verified by digital subtraction angiography (DSA) and computed tomography (CT). Thrombectomy was performed using 6F and 8F Rotarex and 6F, 8F, and 10F Aspirex catheters (Straub Medical AG, Wangs, Switzerland). After intervention, DSA and CT were repeated to evaluate the efficacy of mechanical thrombectomy and to exclude local complications. In addition, pulmonary CT was performed to rule out pulmonary embolism. Finally, all pigs were killed, and iliac veins were dissected to perform macroscopic and histological examination. Thrombus induction was successfully achieved in all animals as verified by DSA and CT. Subsequent thrombectomy lead to incomplete recanalization of the iliac veins with residual thrombi in all cases. However, the use of the 6F and 8F Rotarex catheters caused vessel perforation and retroperitoneal hemorrhage in all cases. Application of the Aspirex device caused one small transmural perforation in a vessel treated with a 10F Aspirex catheter, and this was only seen microscopically. Pulmonary embolism was detected in one animal treated with the Rotarex catheters, whereas no pulmonary emboli were seen in animals treated with the Aspirex catheters. The Aspirex catheter allowed subtotal and safe recanalization of iliac vein thrombosis. In contrast, the use of the Rotarex catheter caused macroscopically obvious vessel perforations in all cases.

Background The receptor activator of NF-κB (RANK), its ligand (RANKL) and osteoprotegerin (OPG) have been reported to play a role in the pathophysiological bone turnover and in the pathogenesis of breast cancer. Based on this we investigated the role of single nucleotide polymorphisms (SNPs) within RANK, RANKL and OPG and their possible association to breast cancer risk. Methods Genomic DNA was obtained from Caucasian participants consisting of 307 female breast cancer patients and 396 gender-matched healthy controls. We studied seven SNPs in the genes of OPG (rs3102735, rs2073618), RANK (rs1805034, rs35211496) and RANKL (rs9533156, rs2277438, rs1054016) using TaqMan genotyping assays. Statistical analyses were performed using the χ2-tests for 2 x 2 and 2 x 3 tables. Results The allelic frequencies (OR: 1.508 CI: 1.127-2.018, p=0.006) and the genotype distribution (p=0.019) of the OPG SNP rs3102735 differed significantly between breast cancer patients and healthy controls. The minor allele C and the corresponding homo- and heterozygous genotypes are more common in breast cancer patients (minor allele C: 18.4% vs. 13.0%; genotype CC: 3.3% vs. 1.3%; genotype CT: 30.3% vs. 23.5%). No significantly changed risk was detected in the other investigated SNPs. Additional analysis showed significant differences when comparing patients with invasive vs. non-invasive tumors (OPG rs2073618) as well as in terms of tumor localization (RANK rs35211496) and body mass index (RANKL rs9533156 and rs1054016). Conclusions This is the first study reporting a significant association of the SNP rs3102735 (OPG) with the susceptibility to develop breast cancer in the Caucasian population.

This paper presents a brief outline of the notion and the system of oncosimulator in conjunction with a high level description of the basics of its core multiscale model simulating clinical tumor response to treatment. The exemplary case of lung cancer preoperatively treated with a combination of chemotherapeutic agents is considered. The core oncosimulator model is based on a primarily top-down, discrete entity - discrete event multiscale simulation approach. The critical process of clinical adaptation of the model by exploiting sets of multiscale data originating from clinical studies/trials is also outlined. Concrete clinical adaptation results are presented. The adaptation process also conveys important aspects of the planned clinical validation procedure since the same type of multiscale data - although not the same data itself- is to be used for clinical validation. By having exploited actual clinical data in conjunction with plausible literature-based values of certain model parameters, a realistic tumor dynamics behavior has been demonstrated. The latter supports the potential of the specific oncosimulator to serve as a personalized treatment optimizer following an eventually successful completion of the clinical adaptation and validation process.

Intravenous fish oil prolongs graft survival in a heterotopic cardiac transplant model (PVG rats (RT 1 c ) × Wistar/Kyoto rats (RT 1 1 )). This study elaborates the impact of the ester group on the immunosuppressive potential of n-3 fatty acids. Twenty per cent fish oil emulsions (glycerol esters, ethyl esters or concentrated ethyl esters; 9g fat/kg body weight/d) were given by continuous intravenous infusion after transplantation until complete rejection. Fish oil glycerol esters prolonged the heart graft survival time to 12.5 vs. 9.5 d (control group infused with a corresponding amount of soybean oil) (p

In order to detect early precancerous changes which do not involve the whole thickness of the epithelium, we used a novel image analysing program based on an IBAS system (Kontron, Germany) to determine nuclear DNA content (NC) as well as average nuclear area (NA) and variation of nuclear area (VA), in the entire epithelium and in three sublayers, parabasal, intermediate, and superficial. DNA aneuploidy was found in only half of the cases classified as 'high-grade' (HG) lesions, comprising carcinoma in situ (CIS) and severe epithelial dysplasias (EDIII), and was chiefly demonstrable in the parabasal third of the epithelium. The other lesions were DNA euploid. HG lesions showed highly significant increases of NA and VA at the lower levels of the epithelium when compared with 'low-grade' (LG) lesions comprising moderate and mild epithelial dysplasias (EDII and EDI). Our data show that the combination of multiparameter image analysis with conventional morphology assists in the objective grading of precancerous lesions and permits the reliable detection of high-grade lesions.

Background Studies in isolated myocytes and isolated heart preparations have suggested that Na + -H + exchange is an important mechanism for myocardial ischemia–reperfusion injury. This study was undertaken to determine whether inhibition of Na + -H + exchange limits infarct size and improves regional systolic shortening in regional ischemia and reperfusion in intact pigs. Methods and Results The left anterior descending coronary artery was occluded in 18 anesthetized and thoracotomized pigs for 45 minutes and then reperfused for 24 hours. As main end points of this study, regional systolic shortening (sonomicrometry) and infarct size (percentage of infarcted to ischemic myocardium) were determined at the end of the experiments. Infarcted myocardium was assessed by histochemistry (tetrazolium stain) and by quantitative histology of one heart slice. The Na + -H + exchange inhibitor Hoe 694 was administered intravenously at a dose of 3 mg/kg in 6 pigs each either 10 minutes before ischemia (group A) or 10 minutes before the onset of reperfusion (group B). Six pigs served as controls (group C). Treatment with Hoe 694 before ischemia decreased histochemical infarct size from 65±18% (control group) to 13±8% ( P P P =.01). These parameters remained unaffected in group B. Conclusions This study clearly demonstrates that Na + -H + exchange is an important mechanism for cell death in myocardial ischemia and reperfusion in intact pigs; thus, inhibition of this exchange system may prove a promising new strategy in the clinical treatment of myocardial ischemia and reperfusion.

Abstract We studied the cellular distribution of angiotensin-converting enzyme (ACE) in the heart related to the cell types involved in left ventricular repair and remodeling before and after myocardial infarction by immunohistochemical techniques using monoclonal and polyclonal antibodies. In noninfarcted myocardium of both human and rat, ACE expression was confined to endothelial cells and subendocardial cell layers of the aortic valve. ACE was prominent in endothelia of small arteries and arterioles, whereas only half the coronary capillaries were immunoreactive and venous vessels were almost completely devoid of the enzyme. In a rat model of myocardial infarction, ACE distribution was determined 1, 3, and 7 days and 2, 3, and 6 weeks after coronary occlusion. Three and 7 days after infarction, endothelial cells of sprouting capillaries and macrophages in the marginal zone of necrosis revealed ACE expression. In both human and rat with the onset of fibrosis, intense staining of the enzyme was found in the marginal zone of the repair tissue. In situ hybridization for collagen type I in the rat revealed that zones with high collagen content had almost no ACE immunoreactivity. Vascular smooth muscle cells and cardiomyocytes revealed no ACE expression throughout the study. We conclude that endothelial cells are the principal source for the expression of ACE after myocardial infarction. The observed induction of ACE with the onset of fibrosis suggests a role of this enzyme that is related to tissue repair and remodeling.

Hintergrund: Am Modell der heterotopen Rattenherzallotransplantation haben wir bereits gezeigt, daβ intravenöse Fettemulsionen in Abhängigkeit von ihrem n-3/n 6-Fettsäureverhältnis abgestuft von immunsuppressiv bis immunneutral wirken. Distelöl (n-3:n-6 = 1:370), Fisch- (7,6:1) und Sojaöl (1:6,5) verlängerten die Trans-plantatüberlebenszeit auf 13,3, 12,3 und 10,4 Tage gegenüber 6,7 Tagen (1:2,1 = 01-kontrollgruppe) bzw. 7,8 Tagen (Kochsalzkontrollgruppe) (p < OOl). In der vor-liegenden Untersuchung wird die Modifikation der Abstoβungszeit mit immunhisto-logischen, zellbiologischen und biochemischen Variablen korreliert. Material und Methodik: 20%ige Emulsionen von Distel-, Fisch-, Sojaöl bzw. einer l:l-Mischung von Distel- und Fischöl (Ölkontrollgruppe) wurden unmittelbar nach der Transplantation kontinuierlich infundiert (9 g Fett/kg Körpergewicht/Tag; n = 10 pro Gruppe). Subpopulationen der infiltrierenden und zirkulierenden immunkompetenten Zellen und die Leukotrien-B4- bzw. -B5-Ausschüttung zirkulierender mononukleärer Zellen wurden zu einem definierten Zeitpunkt (4. postoperativer Tag) analysiert. Ergebnisse: In den beiden Gruppen mit der höchsten Überlebenszeitverlängerung war die Zahl infiltrierender Zellen um bis zu 40% reduziert. In der Fischölgruppe waren zudem die zirkulierenden T-Zellen signifikant vermindert. Leukotrien B4 wurde in alien Gruppen in gleicher Menge, Leukotrien B5 ausschlieβlich in der Fischölgruppe sezerniert. Schluβfolgerungen: Intravenöse Fettemulsionen wirken in Abhängigkeit vom n-3/n 6-Fettsäureverhältnis unterschiedlich immunmodulierend. Sowohl n-6- als auch n-3-Fettsäuren wirken im Überschuβ durch Reduktion der Infiltration und Mobilisation immunkompetenter Zellen immunsuppressiv, n-3-Fettsäuren auch durch Pro-duktion von Leukotrien B5. Sojaöl mit einem ausgewogeneren n-3/n-6-Verhältnis als Distelöl ist signifikant weniger immunsuppressiv, und Fettemulsionen mit einem n-3/n-6-Verhältnis von 1:2 sind immunologisch neutral.

The human neutrophil NB1 glycoprotein (NB1 GP, HNA-2a, CD177) has gained clinical importance for being involved in pulmonary transfusion reactions and immune neutropenias. The NB1 GP shows the unique feature of being expressed only on a neutrophil subpopulation. Recently, we identified splicing defects responsible for an NB1 GP deficiency. In this study, we have investigated the molecular basis of the heterogeneous expression of NB1 GP by separating the 2 neutrophil subpopulations using immunofluorescence followed by single-cell picking or by fluorescence-activated cell sorter. We found a lack of NB1 mRNA in the NB1 GP- cells that remained constant even after granulocyte colony-stimulating factor (G-CSF) administration. Comparing the cDNA sequences of donors with a large (> 60%) and those with a small (< 40%) NB1 GP–expressing subpopulation, we found 6 polymorphisms. Of the 6, 3 were significantly associated with a small NB1 GP–expressing subpopulation, indicating a genetic basis for NB1 GP nonexpression.

Liver biopsy (LB) is a diagnostic procedure to obtain histological diagnosis, grading and staging in individuals with liver diseases. Most commonly LB is performed percutaneously. However, transjugular liver biopsy (TJLB) is considered as an alternative. The aim of this prospective study was to evaluate the diagnostic accuracy of TJLB. TJLB with a semi-automatic Tru-Cut System (18G) was performed in 39 patients with various liver diseases (21 females and 18 males; age range 11–77 years). The number of complete portal tracts (CPTs), lengths and numbers of acquired cores, number of performed cutting steps (passes), and the possibility to obtain histological diagnosis were analyzed. There were no procedure-related complications, and in total 45 procedures were performed. Diagnosis could be established in 88.8 % of all samples, in five cases histology yielded no diagnosis due to an inadequate number of CPTs or sample fragmentation. In average, 4 passes were performed and 4 (range 1–7) cores were obtained. Median core length was 1.1 cm (range 0.4–1.9 cm), median number of CPTs was 7 (range 0–20). Liver fibrosis in general led to a decreased number of CPTs (p

Systemic vasculitides constitute a heterogeneous group of diseases. Autoimmunity mediated by B lymphocytes and their humoral effector mechanisms play a major role in ANCA-associated vasculitis (AAV) as well as in non-ANCA associated primary systemic vasculitides and in the different types of autoimmune connective tissue disorders and rheumatoid arthritis. In order to detect autoantibodies in systemic vasculitides, we screened protein macroarrays of human cDNA expression libraries with sera from patients with ANCA-associated and ANCA-negative primary systemic vasculitides. This approach led to the identification of antibodies against progranulin, a 88 kDA secreted glycoprotein with strong anti-inflammatory activity in the course of disease of giant-cell arteritis/polymyalgia rheumatica (14/65), Takayasu's arteritis (4/13), classical panarteritis nodosa (4/10), Behcet's disease (2/6) and in the course of disease in granulomatosis with polyangiitis (31/75), Churg-Strauss syndrome (7/23) and in microscopic polyangiitis (7/19). In extended screenings the progranulin antibodies were also detected in other autoimmune diseases such as systemic lupus erythematosus (39/91) and rheumatoid arthritis (16/44). Progranulin antibodies were detected only in 1 of 97 healthy controls. Anti-progranulin positive patients with systemic vasculitides, systemic lupus erythematosus or rheumatoid arthritis had significant lower progranulin plasma levels, indicating a neutralizing effect. In light of the anti-inflammatory effects of progranulin, progranulin antibodies might exert pro-inflammatory effects thus contributing to the pathogenesis of the respective autoimmune diseases and might serve as a marker for disease activity. This hypothesis is supported by the fact that a positive progranulin antibody status was associated with active disease in granulomatosis with polyangiitis.

To evaluate the trends in indications for penetrating keratoplasty (PKP) in Homburg/Saar between 2001 and 2010. Retrospective review of 1,200 corneal buttons which underwent PKP that were performed between 2001 and 2010 at the Department of Ophthalmology of Saarland University Hospital, Germany. Indications were classified into eight different groups following histological analysis: keratoconus, Fuchs’ dystrophy, bullous keratopathy, corneal scars, keratitis, regraft, corneal dystrophy other than Fuchs’ dystrophy, and other diagnoses. Two different time periods (between 2001–2005 and between 2006–2010) were analyzed. Keratoconus (25.5 %) was the most common indication for PKP in our study, followed by Fuchs’ dystrophy (21.2 %), bullous keratopathy (14.6 %), corneal scars (14.4 %), keratitis (13.0 %), regraft (7.0 %), non-Fuchs’ dystrophies (2.1 %), and other diagnoses (2.3 %). Comparing the two different time periods, a trend of significantly increasing frequency of keratoconus and Fuchs’ dystrophy, and a decreasing frequency of corneal scars, were found as indications for PKP in our study. Keratoconus was the leading indication for PKP in our series, and had a significantly increasing trend from 2001–2005 to 2006–2010. The percentage of patients with Fuchs’ dystrophy increased, and became the second most common indication for PKP, while the number of PKPs for corneal scars decreased during the last 5 years in our institution.

Background Despite numerous studies, the tumor biology of pleomorphic adenomas, the most common salivary gland tumors, is still not completely defined. In order to identify further candidate genes important for tumor biology of pleomorphic adenomas, extended cytogenetic and molecular analysis are mandatory. Methods We performed a detailed molecular cytogenetic analysis using comparative genomic hybridization (CGH) followed by fluorescence in situ hybridization (FISH) with probes for chromosome X, 16p, 17, and 20 on a large cohort of pleomorphic adenomas (n = 29). Results We could confirm previously described deletions in pleomorphic adenomas affecting 16p, 17, 20q, and 22 by FISH and/or CGH analysis. Moreover, our CGH study revealed novel candidate regions on 8p23.1pter, 9p, 10q25.1q25.3, and 11q24qter in the series of analyzed pleomorphic adenomas. Conclusion Our present study reveals new insights in novel candidate regions implicated in pleomorphic adenoma tumorigenesis which should be considered in further molecular studies. © 2012 Wiley Periodicals, Inc. Head Neck 35: 1431–1438, 2013

Herpes simplex virus type 1 (HSV-1) infections cause typical dermal and mucosal lesions in children and adults. Also complications to the peripheral and central nervous system, pneumonia or hepatitis are well known. However, dissemination to viscera in adults is rare and predominantly observed in immunocompromised patients. Here we describe the case of a 70-year-old male admitted with macrohematuria and signs of acute infection and finally deceasing in a septic shock with multi organ failure 17 days after admission to intensive care unit. No bacterial or fungal infection could be detected during his stay, but only two days before death the patient showed signs of rectal, orolabial and genital herpes infection. The presence of HSV-1 was detected in swabs taken from the lesions, oropharyngeal fluid as well as in plasma. Post-mortem polymerase chain reaction analyses confirmed a disseminated infection with HSV-1 involving various organs and tissues but excluding the central nervous system. Autopsy revealed a predominantly retroperitoneal diffuse large B-cell lymphoma as the suspected origin of immunosuppression underlying herpes simplex dissemination.

Walker and colleagues 1 were the first to define the term “inflammatory aneurysm.” They reported 19 cases and found that these aneurysms were nonbacterial special types of atherosclerotic aneurysms. Inflammatory aneurysms constitute approximately 5% of all infrarenal abdominal aortic aneurysms.2 However, location of these aneurysms in the ascending thoracic aorta is extremely uncommon.3 Connery and colleagues4 first described an isolated inflammatory aneurysm in the ascending thoracic aorta. Typical histologic examination shows signs of chronic inflammation in the adventitia with a marked lymphoplasmacytic cell infiltrate and granulation or fibrous tissue proliferation.5

Introduction: Biocompatibility of implants in humans has been classified as “inert,” “tolerated,” and “bioactive.” In shunt-treated patients, catheter-induced complications account for up to 70% of all hardware failures. Our objective was to study whether foreign body reactions to silicone shunt catheters in subcutaneous tissue and at their distal, intraperitoneal ends leading to occlusion can be reproduced in an animal model.

Background & Aims The insulin-like growth-factor 2 ( IGF2 ) mRNA binding protein p62 is highly expressed in hepatocellular carcinoma tissue. Still, its potential role in liver disease is largely unknown. In this study, we investigated pathophysiological implications of p62 overexpression in mice. Methods We generated mice overexpressing p62 under a LAP-promotor. mRNA expression levels and stability were examined by real-time RT-PCR. Allele-specific expression of Igf2 and H19 was assessed after crossing mice with SD7 animals. The Igf2 downstream mediators pAKT and PTEN were determined by Western blot. Results Hepatic p62 overexpression neither induced inflammatory processes nor liver damage. However, 2.5week old transgenic animals displayed a steatotic phenotype and improved glucose tolerance. p62 overexpression induced the expression of the imprinted genes Igf2 and H19 and their transcriptional regulator Aire (autoimmune regulator). Neither monoallelic expression nor mRNA stability of Igf2 and H19 was affected. Investigating Igf2 downstream signalling pathways showed increased AKT activation and attenuated PTEN expression. Conclusions The induction of a steatotic phenotype implies that p62 plays a role in hepatic pathophysiology.

Angiotensin I-converting enzyme (kininase II, ACE, CD143) availability is a determinant of local angiotensin and kinin concentrations and physiological actions. Limited information is available on ACE synthesis in peripheral vascular beds. We studied the distribution of ACE along the human and rat vascular tree, and determined whether the enzyme was uniformly distributed in all endothelial cells (EC) or if differences occurred among vessels and organs. The distribution of ACE was assessed by using a panel of anti-human ACE monoclonal antibodies and serial sections of the entire vascular tree of humans. Comparison was made with other EC markers. EC of small muscular arteries and arterioles displayed high ACE immunoreactivity in all organs studied except the kidney, while EC of large arteries and of veins were poorly reactive or completely negative. Only 20% on average of capillary EC in each organ, including the heart, stained for ACE, with the remarkable exception of the lung and kidney. In the lung all capillary EC were labeled intensively for ACE, whereas in the kidney the entire vasculature was devoid of detectable enzyme. In contrast to the man, the rat showed homogeneous endothelial expression of ACE in all large and middle-sized arteries, and in veins, but in renal vessels ACE expression was reduced. This study documents a vessel, organ and species specific pattern of distribution of endothelial ACE. The markedly reduced ACE content of the renal vasculature may protect the renal circulation against excess angiotensin II formation and kinin depletion, and maintain high renal blood flow.

Background The apoE-/-/LDL-/- double knockout mice are bearing considerable structural homology to human atherosclerosis. We hypothesized, that advanced lesion formation in the renal artery is associated with kidney alterations in these mice. Methods Kidneys from apoE-/-/LDL-/- double knockout mice at the age of 80 weeks (n = 6) and C57/BL control mice (n = 5) were infused with Microfil, harvested and scanned with micro-CT (12 μm cubic voxels) and Nano-CT (900 nm cubic voxels). We quantitated the total vascular volume using micro-CT. Number and cross-sectional area (μm2) of glomeruli were measured using histology. Results At the age of 80 weeks, the renal total vascular volume fraction decreased significantly (p < 0.001) compared to controls. Moreover, the renal artery showed advanced atherosclerotic lesions with adventitial Vasa vasorum neovascularization. Perivascular inflammation was present in kidneys of apoE-/-/LDL-/- double knockout mice, predominantly involved are plasma cells and leucocytes. Glomeruli cross-sectional area (9959 ± 1083 μm2) and number (24.8 ± 4.5) increased in apoE-/-/LDL-/- double knockout mice compared to controls (3533 ± 398 μm2; 17.6 ± 3, respectively), whereas 41% of the total number of glomeruli showed evidence for lipoprotein associated glomerulopathy (LPG). Moreover, immunohistochemistry demonstrated capillary aneurysms of the glomeruli filled with factor 8 containing emboli. Conclusion The reduced intra-renal total vascular volume is associated with systemic atherosclerosis and glomeruli alterations in the apoE-/-/LDL-/- double knockout mouse model.

Structural data about the human lung fine structure are mainly based on stereological methods applied to serial sections. As these methods utilize 2D images, which are often not contiguous, they suffer from inaccuracies which are overcome by analysis of 3D micro-CT images of the never-sectioned specimen. The purpose of our study was to generate a complete data set of the intact three-dimensional architecture of the human acinus using high-resolution synchrotron-based micro-CT (synMCT). A human lung was inflation-fixed by formaldehyde ventilation and then scanned in a 64-slice CT over its apex to base extent. Lung samples (8-mm diameter, 10-mm height, N = 12) were punched out, stained with osmium tetroxide, and scanned using synMCT at (4 {micro}m){sup 3} voxel size. The lung functional unit (acinus, N = 8) was segmented from the 3D tomographic image using an automated tree-analysis software program. Morphometric data of the lung were analyzed by ANOVA. Intra-acinar airways branching occurred over 11 generations. The mean acinar volume was 131.3 {+-} 29.2 mm{sup 3} (range, 92.5-171.3 mm{sup 3}) and the mean acinar surface was calculated with 1012 {+-} 26 cm{sup 2}. The airway internal diameter (starting from the bronchiolus terminalis) decreases distally from 0.66 {+-} 0.04 mm tomore » 0.34 {+-} 0.06 mm (P < 0.001) and remains constant after the seventh generation (P < 0.5). The length of each generation ranges between 0.52 and 0.93 mm and did not show significant differences between the second and eleventh generation. The branching angle between daughter branches varies between 113-degree and 134-degree without significant differences between the generations (P < 0.3). This study demonstrates the feasibility of quantitating the 3D structure of the human acinus at the spatial resolution readily achievable using synMCT.« less

Summary Neurogenic neoplasms account for approximately 1% of salivary gland tumors. A 45-year-old woman presented with a slowly growing unilateral parotid gland mass that was excised completely by lateral parotidectomy. No recurrence was detectable 6 months after surgery. The resection specimen contained a 1.7 × 1.2 × 1–cm whitish nodular lesion that was firm, well circumscribed, and completely surrounded by compressed normal glandular tissue. The tumor was composed of spindle cells with bipolar tapering, wavy nuclei, and palely stained cytoplasm. The tumor cells were arranged in storiform, lamellar, and whorled patterns. Nuclear atypia, necrosis, and significant mitotic activity were absent. Immunohistochemistry showed expression of epithelial membrane antigen, glucose transporter 1, claudin-1, and collagen IV. All other lineage-specific markers including protein S100 were negative in the tumor cells. To our knowledge, this case represents the first report of soft tissue perineurioma in the salivary gland. Lack of previous reports suggests underrecognition of this tumor entity at this unusual anatomical site.

Ziele: To evaluate the distribution of vasa vasorum (VV) in advanced atherosclerotic lesions (calcified, fibrotic or hemorrhaged) in the aortic wall of apoE-/-/LDL-/- mice in relation to aortic luminal and plaque cross-sectional surface area using high-resolution nano-CT Methode: Aortas from male apoE-/-/LDL-/- mice (n=7) at the age of 80 weeks were infused in situ with contrast agent, harvested for scanning with Nano-CT (NCT) and followed by sectioning for histology. The spatial distribution of Vasa Vasorum [number and surface/cross-section (mm2)] was compared to aortic luminal cross-sectional area in the ascending aorta, aortic arch and descending aorta. Results were complemented with co-localized histology. Ergebnis: The number and cross-sectional area of VV in relation to plaque showed significant differences depending on whether it was a fibrotic, hemorrhaged or calcified plaque (6.8±1.9 vs. 6.9±2.4 vs. 1.4±1.3 respectively, p

It is unknown whether transforming growth factor beta1 (TGF-beta1) signaling uniformly participates in fibrogenic chronic liver diseases, irrespective of the underlying origin, or if other cytokines such as interleukin (IL)-13 share in fibrogenesis (e.g., due to regulatory effects on type I pro-collagen expression). TGF-beta1 signaling events were scored in 396 liver tissue samples from patients with diverse chronic liver diseases, including hepatitis B virus (HBV), hepatitis C virus (HCV), Schistosoma japonicum infection, and steatosis/steatohepatitis. Phospho-Smad2 staining correlated significantly with fibrotic stage in patients with HBV infection (n = 112, P0.001) and steatosis/steatohepatitis (n = 120, P0.01), but not in patients with HCV infection (n = 77, P0.05). In tissue with HBx protein expression, phospho-Smad2 was detectable, suggesting a functional link between viral protein expression and TGF-beta1 signaling. For IL-13, immunostaining correlated with fibrotic stage in patients with HCV infection and steatosis/steatohepatitis. IL-13 protein was more abundant in liver tissue lysates from three HCV patients compared with controls, as were IL-13 serum levels in 68 patients with chronic HCV infection compared with 20 healthy volunteers (72.87 +/- 26.38 versus 45.41 +/- 3.73, P0.001). Immunohistochemistry results suggest that IL-13-mediated liver fibrogenesis may take place in the absence of phospho-signal transducer and activator of transcription protein 6 signaling. In a subgroup of patients with advanced liver fibrosis (stageor =3), neither TGF-beta nor IL-13 signaling was detectable.Depending on the cause of liver damage, a predominance of TGF-beta or IL-13 signaling is found. TGF-beta1 predominance is detected in HBV-related liver fibrogenesis and IL-13 predominance in chronic HCV infection. In some instances, the underlying fibrogenic mediator remains enigmatic.

OBJECTIVE This study evaluated the influence of voxel size on its ability to discriminate calcium from iron deposits in ex vivo coronary arteries. METHODS Postmortem human coronary arteries underwent multislice computed tomographic scan at (600-microm) voxel size to provide an index of computed tomography (CT) image noise and synchrotron-based micro-CT at (4-microm) voxel size to provide data for generating a range of voxel sizes 4 to (600-microm) after grayscale noise was added to the projection images before reconstruction so as to mimic the effect of retaining the same radiation exposure involved in the multislice computed tomographic scan. RESULTS At voxel sizes of (20-microm) or smaller, iron deposits could be identified based on CT grayscale value. Voxels of (100-microm) or larger cannot resolve nor distinguish iron deposits from calcifications by virtue of CT grayscale value. CONCLUSIONS Clinical CT scanners cannot be expected to discriminate iron deposits from calcifications by their CT value alone in the arterial wall.

The ContraCancrum project aims at developing a composite multilevel platform for simulating malignant tumor development and tumor and normal tissue response to therapeutic modalities and treatment schedules. The project aims at having an impact primarily in (a) the better under-standing of the natural phenomenon of cancer at different levels of biocomplexity and most importantly (b) the disease treatment optimization procedure in the patient’s individualized context by simulating the response to various therapeutic regimens. Fundamental biological mechanisms involved in tumor development and tumor and normal tissue treatment response such as metabolism, cell cycle, tissue mechanics, cell survival following treatment etc. are modeled also addressing stem cells in the context of both tumor and normal tissue behavior. The simulators exploit several discrete and continuous mathematics methods such as cellular automata, the generic Monte Carlo technique, finite elements, differential equations, novel dedicated algorithms etc. The predictions of the simulators rely on the imaging, histopathological, molecular and clinical data of the patient. ContraCancrum deploys two important clinical studies for validating the models, one on lung cancer and one on gliomas. The crucial validation work is based on comparing the multi-level therapy simulation predictions with multi-level patient data, acquired before and after therapy. ContraCancrum aims to pave the way for translating clinically validated multilevel cancer models into clinical practice.