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Um die Ursachen fur das Auftreten invasiver Zervixkarzinome trotz zytologischer Vorsorgeuntersuchung in Karnten genauer zu ermitteln, wurden in den Jahren 2000–2002 alle 132 Patientinnen mit diagnostiziertem Zervixkarzinom erfasst und einer Recherche im Hinblick auf das Vorliegen von gynakologisch-zytologischen Abstrichen innerhalb der letzten 5 Jahre vor der Diagnosestellung des Karzinoms unterzogen. Bei 50% der Patientinnen mit einem Zervixkarzinom aus den Jahren 2000–2002 konnten in den 5 Jahren davor keine gynakologisch-zytologischen Abstriche ermittelt werden. Im Jahr 2002 wurden 53 Patientinnen mit Zervixkarzinom gemeldet, und 78 erfassbare Abstriche wurden reevaluiert. 49% aller primar negativ befundeten Abstriche wurden in der Reevaluation als positiv (≥Pap III) eingestuft. 92% aller als „richtig“-negativ befundeten Abstriche wiesen in der Reevaluation Qualitatsmangel des Abstrichs auf. Die Befundervariabilitat (Kappa-Statistik) war beim Vergleich der Ergebnisse der Erstbefunde im Screening und der zusammengefassten Ergebnisse der Nachbefunder masig gut, ebenso die Variabilitat innerhalb der Nachbefunder.

Cervical cancer screening developed rapidly during the 1970s. Today, approximately 1.5 million smears are taken annually, so 50% of the target population are screened every year, 30% are cytologically underserved (24% never had a smears, 6% only once). This figure correlates with the fact that there are still 30% deaths from cervical cancer compared with 1960. Since 1998 a voluntary quality assurance programme was introduced by the Austrian Society of Cytology, based on comparison of results reported from participating laboratories.

Summary Aim: The goal of this study was to assess the accuracy and limitations of ultrasound guided fine-needle aspiration biopsy (ug-FNAB) of solitary thyroid nodules. Methods: The ug-FNAB results of 538 patients with solitary thyroid nodules, who afterwards underwent thyroid surgery, were compared retrospectively with the histology. Patients with multinodular goiter were excluded from the study. Ug-FNAB was performed on growing and/or hypoechoic and/or hypofunctional nodules. The ug-FNAB results were grouped as follows: group 1: malignant (n = 44); group 2: malignancy cannot be ruled out (n = 173); group 3: non-malignant (n = 296), group 4: inadequate (n = 25). Results: When the cytological results of group 1 and group 2 were interpreted os being malignant and those of group 3 as being benign, sensitivity, specificity and accuracy of ug-FNAB were 96.7%, 65.8% and 69.5% respectively. The 62 thyroid carcinomas (TO biopsied presented in 59 cases a suspicious or malignant cytology (95.2%). The smallest TC diagnosed by ug-FNAB had a diameter of 0.5cm and 36.4% of all papillary TC ≤ 1 cm displayed stage pT4. The histology verified a TC in 18 cases out of the 173 ug-FNABs in group 2. Non-malignant ug-FNABs were confirmed by histology in 294 patients (99.3%) in group 3. In 4.65% of the ug-FNABs inadequate material was aspirated. Conclusion: Nodules with non-suspicious ug-FNAB results can be safely followed-up by sonography, as the cytological diagnoses were verified in more than 99% by histology. Papillary TC can be diagnosed with ug-FNAB very accurately. As stage pT4 was present in more than one third of patients with papillary TC ≤ T cm, ug-FNAB is also recommended for thyroid nodules 0.5-1 cm in diameter located adjacent to the thyroid capsule. However, microfollicular proliferations remain the limitation of ug-FNAB, as the cytology cannot distinguish between benign adenoma and follicular TC.

The aim of this study was to determine the value, advantages and limitations of ultrasound-guided fine-needle aspiration biopsy (US-FNAB) in an endemic goitre area. US-FNAB was performed on all outpatients who presented with hypoechoic and/or hypofunctional and/or growing nodules. A total of 4518 US-FNABs were performed and 718 patients from this series underwent surgery. Cytological results of the primarily performed US-FNAB of these patients were compared retrospectively with the histological results. US-FNAB results were grouped as (1) non-malignant (n = 303), (2) non-malignant follicular proliferation (n = 177), (3) malignancy cannot be ruled out (n = 133), (4) malignant (n = 61), (5) inadequate (n = 34), and (6) sampling error; biopsy of a non-malignant nodule (n = 10). Nodules as small as 5 mm in diameter could be biopsied, gaining representative material. US-FNAB found a malignant or suspicious cytology in 65 out of 87 cases with malignant histology (74.71%). Diagnosis of early tumour stages was often possible: 12 of 18 thyroid carcinomas biopsied and smaller than 10 mm in diameter had malignant or suspicious cytology (groups 3 and 4). US-FNAB was performed incorrectly within non-malignant nodules in ten patients (1.39%) with multinodular goitre (ten papillary carcinomas, nine smaller than 10 mm). Regarding the cytology of groups 1 and 2 as benign and those of groups 3 and 4 as malignant, US-FNAB performance was as follows: sensitivity 87.84%, specificity 78.50%, negative predictive values 98.13%, positive predictive values 33.51% and accuracy 79.53%. Biopsies with inadequate material were obtained in 4.73% of all biopsies. No major adverse effects occurred. Re-biopsies in 61 cases did not alter the cytological outcome in those cases where adequate material was obtained. US-FNAB is a valuable method in the pre-operative assessment of thyroid nodules in order to select patients for surgery, as malignancy can often be detected even in early tumour stages. However, even with ultrasonographic guidance, the minimal tumour size detectable by US-FNAB is around 5 mm. The cytological interpretation in cases with regression and microfollicular proliferation also sets limits on the method. However, patients with non-malignant cytologies can be followed up safely by sonography due to the high NPV of US-FNAB as long as thyroid nodules do not become larger. Re-biopsies seem to be of limited value as long as adequate material was obtained by US-FNAB.

To evaluate the reasons for the occurrence of invasive cervical cancer in Carinthia despite cytological screening, all 132 patients diagnosed with cervical cancer in the years 2000-2002 were recorded and all gynecological cytological smears made within the 5 years prior to the diagnosis of cancer were reevaluated.Within the 5 years prior to diagnosis, no gynaecological cytological smear was found for 50% of the patients diagnosed with cervical cancer in the years 2000-2002. In the year 2002, a total of 53 patients were reported to have cervical cancer and 78 smears were reevaluated. Of all the smears primarily diagnosed as negative, 49% were found to be positive (or =Pap III) after reevaluation and 92% of all smears "correctly" diagnosed as negative showed quality deficiencies. The interobserver variability (kappa-statistics) showed a moderate value when the primary screening results were compared with the reevaluation. The interobserver variability within the group of reevaluators was also moderate.

ZusammenfassungEs wird über eine 73jährige Patientin mit dem sehr seltenen Fall eines Karzinoms berichtet, das sich in einer Ductus thyreoglossus-Zyste innerhalb einer Struma diffusa et nodosa entwickelt hatte. Zusätzlich bestand eine Immunthyreopathie Typ Basedow. Sonographische, röntgenologische, szintigraphische und histologische Befunde werden vorgestellt.

The goal of this study was to assess the accuracy and limitations of ultrasound guided fine-needle aspiration biopsy (ug-FNAB) of solitary thyroid nodules.The ug-FNAB results of 538 patients with solitary thyroid nodules, who afterwards underwent thyroid surgery, were compared retrospectively with the histology. Patients with multinodular goiter were excluded from the study. Ug-FNAB was performed on growing and/or hypoechoic and/or hypofunctional nodules. The ug-FNAB results were grouped as follows: group 1: malignant (n = 44); group 2: malignancy cannot be ruled out (n = 173); group 3: non-malignant (n = 296), group 4: inadequate (n = 25).When the cytological results of group 1 and group 2 were interpreted as being malignant and those of group 3 as being benign, sensitivity, specificity and accuracy of ug-FNAB were 96.7%, 65.8% and 69.5% respectively. The 62 thyroid carcinomas (TC) biopsied presented in 59 cases a suspicious or malignant cytology (95.2%). The smallest TC diagnosed by ug-FNAB had a diameter of 0.5 cm and 36.4% of all papillary TCor = 1 cm displayed stage pT4. The histology verified a TC in 18 cases out of the 173 ug-FNABs in group 2. Non-malignant ug-FNABs were confirmed by histology in 294 patients (99.3%) in group 3. In 4.65% of the ug-FNABs inadequate material was aspirated.Nodules with non-suspicious ug-FNAB results can be safely followed-up by sonography, as the cytological diagnoses were verified in more than 99% by histology. Papillary TC can be diagnosed with ug-FNAB very accurately. As stage pT4 was present in more than one third of patients with papillary TCor = 1 cm, ug-FNAB is also recommended for thyroid nodules 0.5-1 cm in diameter located adjacent to the thyroid capsule. However, microfollicular proliferations remain the limitation of ug-FNAB, as the cytology cannot distinguish between benign adenoma and follicular TC.

In the follow up of differentiated thyroid carcinoma (DTC) several scintigraphic methods are used in addition to the serum thyroglobulin and ultrasonography of the neck. Iodine-131 whole body scintigraphy (WBS), which is performed since many years, is able to detect iodine positive recurrence, lymph node metastases and distant metastases in a very specific way. However, the problem of I-131 WBS is the fact that only 67% of metastases from DTC accumulate iodine. Therefore non specific radionuclides like TI-201 or tracers such as Tc-99m Sestamibi or Tc-99m Tetrofosmin and new metabolic tracers like F-18 FDG were introduced in the diagnostic work up to detect iodine negative metastases as well. This study describes the comparison of different tracers in 35 patients with elevated thyroglobulin and suspicion of metastatic disease or already known metastases from DTC.

In patients with suspicious breast lesions, scintimammography (SM) using Tc-99m tetrofosmin (TETRO) and hydroxymethylene-diphosphonate (HDP) and Gd-DTPA MRI were performed. For both TETRO and MRI sensitivity was 93% compared to 71% for HDP; specificity was lower for TETRO and HDP (71%) than for MRI (86%). This preliminary study shows that, in contrast to HDP, TETRO SM and Gd-DTPA MRI are of additional diagnostic value in diagnosing breast cancer.

We report a case of a 48-yr-old woman who underwent surgery because of papillary oxyphilic thyroid carcinoma pT3. After total thyroidectomy, we administered 2960 MBq (131)I for ablation of the residual tissue. initial follow-up visits showed no clinical, radiological or scintigraphic evidence of residual or metastatic thyroid tissue. Serum thyroglobulin levels (Tg) and (131)I whole-body scintigraphy were negative. Three years after thyroidectomy, the patient experienced seizures, and as a consequence a brain tumor was removed. It was an undetected metastasis of the primary thyroid carcinoma. Histological examinations showed that neither the primary tumor nor the metastasis produced any Tg. With this fact in mind and the knowledge of negative (131)I whole-body scans we had to concentrate on radiological (CT and MRI scans) and nonspecific scintigraphic methods such as 201TI whole-body scintigraphy in our management of the patient. Further follow-up demonstrated multiple metastasis by 201TI whole-body scan (mediastinum, bones and soft tissue), and most of them have been removed by surgery. This case report demonstrates that, in addition to (131)I whole-body scans and measurement of serum Tg, the use of nonspecific tracers like 201TI is important to detect (131)I and/or Tg negative metastases.

The uptake of 99mTc-tetrofosmin in enlarged lymph nodes, of the lung hilus, in the case of sarcoidosis Stage I (histopathologically confirmed by mediastinoscopic biopsy) is demonstrated. On a routine chest radiograph of a 78-yr-old woman, hilar lymphadenopathy was first detected. In the following mammography, disseminated micro calcifications were found in the left breast and a 99mTc-tetrofosmin study was performed for detection of breast cancer. Scintigraphy using 99mTc-tetrofosmin showed clear uptake in the hilar lymph nodes, but not in the left breast. The 99mTc-tetrofosmin uptake in the hilar lymph nodes was due to sarcoidosis confirmed by histology. Therefore, 99mTc-tetrofosmin scintigraphy may be useful in patients with suspected sarcoidosis, especially in Stage I.

In a 42-year-old women a recurrent anaplastic ependymoma of the frontal lobe developed intracerebral metastases and, after implantation of an atrioventricular shunt, also metastasized to the lung, pleural cavity and mediastinal lymph nodes. In routinely stained smear preparations of the pleural effusion sediment, the typical cytologic features of an anaplastic ependymoma were found--papilliform cell clusters with acellular axes ("pseudorosettes"), incomplete true ependymal rosettes, and epithelioid and tanycytoid tumor cells with signs of anaplasia. A peculiar matrix material was a characteristic feature of this case and, to our knowledge, was not reported previously. It consisted of refringent, fibrillar strands composed of tightly packed, roundish to rodshaped granules. This matrix seemed to be associated with the cytoplasmic processes of the tumor cells.

A 73 y old female was found to suffer from a squamous cell carcinoma with a focal thymus-like differentiation in a thyroglossal duct cyst. A neoplasma of this type in a goitre with Graves' disease is extremely rare. Sonographic, radiological, scintigraphic and microscopic findings are discussed.

Histologic investigation of liver tissue obtained by percutaneous needle biopsy remains the mainstay in the diagnosis of liver disease. Impaired blood clotting or thrombocytopenia precludes percutaneous liver biopsy for the danger of sometimes life-threatening hemorrhage as the most serious complication. The feasibility of plugging the intrahepatic needle track with fibrin sealant to minimize the risk of bleeding has been shown. The advantage of the method presented here is that biopsy and sealing can be performed as a one-step procedure, thus shortening the time necessary and minimizing the risk of complications. We considered laparoscopic control most reliable to optimize handling of the double-channel needles and to assess the reliability of fibrin plugging for the “blind” percutaneous approach. In 27 patients a modified Menghini needle and in 10 patients a modified Trucut needle were tested. In our experience the handling of the modified Trucut needle proved easier. There was, however, no significant difference in the length of the biopsy core obtained with the two needles (16 ± 8.6 mm Menghini needle and 15.75 ± 7.1 mm Trucut needle), resulting from the larger diameter of the Menghini needle (1.8). The thin trickle of bleeding observed around the plug in a few patients was without clinical significance, reflecting the prolonged coagulation time in these patients. In summary, in the hands of an experienced investigator the combined biopsy-plugging device is a safe and reliable tool for obtaining liver tissue in patients with impaired blood coagulation. As a one-step procedure it can be performed quickly, thus increasing safety by diminishing the need for patient cooperation.

An extraordinary rare case of implant metastasis at the laparoscopic working port, two years after an apparent uncomplicated laparoscopic cholecystectomy is presented. The histological reexamination of the gallbladder with the detection of a minimal carcinomatous lymphangiosis in newly prepared sections, the exclusion of other primary tumors and also the tumor-free interval of six months until now, after radical excision of the abdominal wall, are strongly suggestive for an intraoperative tumor cell dissemination into the abdominal wall by a histologically unrecognized carcinoma of the gallbladder.

A systematic immunohistochemical study of different stages of atherosclerosis in human aortas was performed using several antibodies. Because oxidation of lipoproteins could be a key event in atherogenesis, an antibody against apolipoprotein B (apoB) from low-density lipoprotein (LDL) modified with the lipid peroxidation-specific aldehyde, 4-hydroxynonenal (4-HNE) (anti-4-HNE-apoB), was raised in rabbits. This antibody recognizing 4-HNE protein adducts was used in concert with an antibody to apo(a) from lipoprotein(a), considered also potentially atherogenic, as well as with an antibody and a monoclonal antibody (mAb) to apoB. Autopsy material from 12 corpses was investigated. The immunohistochemical investigation by the alkaline-phosphatase technique included control specimens regarding postmortem artifacts by autolysis and oxidation. The results from six specimens from five corpses are presented. A positive staining with the antibody to apoB but not with anti-4-HNE-apoB was seen in the normal intima. The thickened intima of early, transitional, and advanced atherosclerotic lesions and atheromata showed a predominantly extracellular staining with all antibodies and the applied mAb. To test the specificity of the staining, antibodies preadsorbed by the appropriate antigens and nonimmune sera were used, giving negative results. These findings indicated a colocalization of epitopes derived from lipid peroxidation of polyunsaturated fatty acids and epitopes specific for apoB and apo(a) during atherogenesis in humans.

Impaired blood clotting precludes percutaneous liver biopsy for histologic examination of liver tissue. The transjugular or laparoscopic approach are ways to reduce the risk of bleeding. These techniques, however, are laborious and confined to specialized centers. Methods of plugging the needle track with sealant presented so far are hampered by the need to leave either the cannula in situ or the need for a second percutaneous approach for the application of the sealant. We have tested a combined plugging-biopsy device allowing to perform the biopsy as a one-step procedure in patients with impaired clotting under laparoscopic vision control. In 37 patients either a modified Trucut or Menghini needle was tested. Handling of the Trucut needle proved easier. Tissue yield was satisfactory with both needles. Only one episode of bleeding was observed with a prototype Menghini needle. We conclude, that the combined plugging-biopsy device is a safe and reliable tool for obtaining liver tissue in patients with impaired blood coagulation.

The angiogenic and fibrogenic activities of fibrin sealant (Tisseel) suggest that it could enhance osteoinductivity during the first days after implantation. To test this hypothesis, the osteoinductivity of sealed and unsealed bone matrix gelatin was tested by implantation into rat abdominal wall muscle pouches for two to ten days. A sponge of collagen with or without fibrin sealant was used for control. Alkaline phosphatase (ALPase) was employed to quantify osteoinduction, and myeloperoxidase (MPO) was used to determine the inflammatory reaction. Plain histologic analysis of bone matrix gelatin implants at Days 2, 4, and 10 did not demonstrate any significant differences between sealed and nonsealed implants. Histologic analysis at Day 21 confirmed the osteoinductivity of the implant. The combination of bone matrix gelatin and fibrin sealant induced an early significant increase in ALPase activity, which could be interpreted as early induction of histologically undetectable osteogenesis. A simultaneous increase in the level of MPO suggested that the bone matrix gelatin/fibrin sealant-induced ALPase reaction might be caused by an inflammatory process, but neutrophil counting did not correlate to the MPO data. Morphometry, however, seems to be insufficient to assay a heterogenous histologic distribution of singular cells. It is proposed that the ALP/MPO ratio may be used as an inflammation-independent marker of osteogenesis. This study does not support the notion of an osteoinductive potency of fibrin sealant per se; however, it is clear that fibrin sealant does not impair bone matrix gelatin-dependent osteoinduction.

Leukocytes, in particular polymorphonuclear neutrophils (PMNs), release important mediators of microvascular injury seen after ischemia/reperfusion, hemorrhage, and sepsis. Adherence of PMNs to the endothelium is thought to be crucial in PMN-mediated injury and results in an increase in permeability and edema. Adhesion is also an in tegral part of the beneficial role of leukocytes during migration, phagocytosis and bacterial killing.

The liver in acute shock is usually enlarged and may be purplish in congestive heart failure (Sherlock 1987) or pale in hypovolemic-traumatic shock. Sometimes the surface of the shocked liver shows a patchy appearance with anemic spots because of circumscribed ischemic areas. The edges of the liver are rounded and because of the interstitial edema the liver weight is increased. Spotty subcapsular hemorrhages may also occur, especially in septic shock, with consumption of coagulation factors resulting in hemorrhagic diathesis. The cut surface of the liver is grayish and pale in pure hypovolemic shock or reveals distended hepatic veins in congestive heart failure. In addition the cut surface may show anemic spots or hemorrhagic patches. Pale and dry areas of yellow color represent necrotic foci; however, such macroscopically detectable necrotic foci are usually rare since they must become quite large to be detectable in this way.

This immunohistochemical study was designed to investigate the possible contribution to and topographical distribution of some important cytokines, such as tumour necrosis factor alpha (TNF alpha) and interleukins, in acute alcoholic hepatitis. The well-known inductive capacity of these cytokines with respect to the expression and/or up-regulation of adhesion molecules, such as intercellular adhesion molecule-1 (ICAM-1) and endothelial leukocyte adhesion molecule-1 (ELAM-1), was a further point to be studied. Moreover, the proposed induction of adhesion molecules might also be associated with the activation and attraction of a special population of inflammatory cells characteristic for alcoholic hepatitis. Frozen liver samples from patients who died with signs of acute alcoholic hepatitis were evaluated using the alkaline phosphatase anti-alkaline phosphatase immunostaining technique and also single and double indirect immunofluorescence. In acute alcoholic hepatitis TNF alpha could be detected predominantly in ballooned hepatocytes, which often contained alcoholic hyalin (Mallory bodies). Moreover, TNF alpha showed a co-distribution with ICAM-1 expressed in the membranes of hepatocytes and with the occurrence of CD11b positive polymorphonuclear leukocytes (neutrophils) suggesting a possible major role of the beta 2-integrin Mac-1 as a ligand for ICAM-1. No induction of ELAM-1 could be found. In alcoholic hepatitis cytokines may be responsible for the induction of the adhesion molecule ICAM-1 on hepatocytic membranes and activate a defined population of inflammatory cells, thus contributing to the characteristic histological picture of acute alcoholic hepatitis with its concentration of neutrophils especially in areas with ballooned Mallory body-containing hepatocytes. Our results are in line with clinical findings showing high levels of TNF alpha and interleukin-1 in sera of patients with alcoholic hepatitis and with the already reported expression of ICAM-1 on hepatocytes.

The development of organ failure may occur within the first days after trauma (early organ failure) or after one or two weeks (late organ failure) in ICU patients. The reaction that leads to organ damage and organ failure is a generalized inflammatory event [1]. This inflammation may be non-bacterial in the early stage of organ failure, and be aggravated by the influence of bacterial factors during the development of late organ failure. The non-bacterial inflammatory reaction is induced by the initial injury and is usually based on three factors: tissue ischemia/hypoxia, reperfusion events with generation of oxygen radicals; stimulation of phagocytes by complement split products or other humoral inflammatory factors; and the interaction with endothelial cells. This chapter focuses on PMN/endothelial activation with special emphasis on the PMN-endothelial interaction. The results of these events will be discussed in the trauma and the sepsis situation.

Summary A method has been established for storage and preservation of cytological specimens in liquid nitrogen and further processing for immunocytochemistry as smears prepared from thawed cells or cryo-sections of frozen cell pellets. For the experiments cultured cells of a T-lymphoblastic leukemia cell line (ATCC CCL 119) and blood cells ofthe buffy coat ofhealthy humans were treated with a cryo-solution (fetal calf serum +5 % dimethylsulfoxid) and after freezing stored in liquid nitrogen. Alternatively, cells preincubated with cryo-solution followed by suspension in fetal calf serum without cryo-additive were frozen and stored in liquid nitrogen for the production of cryo-sections. Indirect immunofluorescence and alkaline phosphatase — antialkaline phosphatase based immunoreactions were performed for the decoration of various surface antigens with a panel of monoclonal antibodies. All immunoreactions were repeated at least three times and the stored cell preparations were investigated after different periods of storage (up to four months). The immunoreactions of fresh cells in suspension (which were used as controls) were comparable with those of cryopreserved cells, e.g. cells on smears after thawing and on cryo-sections of cell pellets. The strongest immunoreactions were achieved on fixed cryo-sections. The maintenance of cell morphology of smears from cryopreserved cells was slightly better than of cells from cryo-sections. In our hands the preparation of cell pellets, which are suitable for the storage in liquid nitrogen and the production of cryosections, is a very useful method for immunocytochemical investigations of cytological specimens especially in situations where immunoreactions cannot be performed on fresh material. Moreover, it provides a possibility for repetition or modification of the immunohistochemical reactions for the same cell population.

Severe ethanol-induced liver damage is characterized by fibrous dissociation of liver cell plates leading to many apparently isolated hepatocytes. Three-dimensional reconstruction, however, revealed hepatocytes that were surrounded by connective tissue as endpoints of "parenchymal pillars" or in association with liver cell plates and bile ductules. Double immunofluorescence studies displayed the expression of cytokeratin (CK) 7 in bile ducts, including bile ductules, but also in some hepatocytes still organized in liver cell plates. The other bile duct, typical CK, namely CK 19, was only detectable in few hepatocytes. However, the expression of CK 7 and/or CK 19 was less frequent in hepatocytes that were closely associated with bile ductules. CK 7 and CK 19 were also found in some, but not all, Mallory bodies. These observations indicate that the expression of these two CKs is neither related to a transformation of hepatocytes to bile duct-like structures ("ductal metaplasia") nor to the formation of Mallory bodies. Furthermore, double immunofluorescence studies revealed small groups of hepatocytes and bile ductules that were encircled by basement membrane material, thus suggesting the formation of "secretory units."

Baboons were subjected to septic or traumatic/hypovolemic shock and their tissues were examined for the de novo expression of endothelial leukocyte adhesion molecule 1 (ELAM-1), using immunohistochemical techniques. In animals with septic shock induced with live Escherichia coli, there was widespread expression of ELAM-1, recognized by monoclonal antibodies H4/18 or ENA-1 in most tissues examined with strong staining in the lung, liver, and kidneys. Endothelial leukocyte adhesion molecule 1 expression was evident in capillaries, venules, small veins, arterioles, and arteries. In contrast, baboons with traumatic/hypovolemic shock had minimal levels of focal ELAM expression in all organs studied. Similarly evidence of neutrophil activation, measured by granulocyte elastase levels in the plasma was much more pronounced in animals with septic shock. The study documents that lipopolysaccharide (LPS)- and cytokine-induced endothelial activation occurs in vivo in septic shock. Much higher levels of ELAM-1 expression and plasma granulocyte-elastase titer in septic shock, as contrasted with traumatic/hypovolemic shock, are consistent with the higher levels of circulating tumor necrosis factor, other cytokines, and LPS in sepsis.

In posttraumatic organ failure we distinguish two different phases -early and late organ failure —, which by definition are caused by non-bacterial (early) and bacterial products (late) induced generalized inflammation. Organ failure is the final result of the action of inflammatory mediators, which cause a permeability increase as well as cellular edema.

Im Zuge eines Polytraumas kommt es neben der direkten Gewebsschadigung zur Ausbildung eines hypovolamischen Schockzustands mit einer Minderperfusion von lebenswichtigen Organen. Daruber hinaus werden weniger wichtige Korperteile von der Durchblutung weitgehen ausgeschaltet, wie z. B. die quergestreifte Muskulatur oder die Haut.

In 11 testes of different developmental stages (from 10-week-old embryos to adult) the cytokeratin and vimentin expression patterns of rete testis and epididymis were investigated immunohistochemically in formaldehyde-fixed paraffin-embedded material. In addition, immunofluorescence microscopy including double immunofluorescence was performed on frozen sections of 3 of these 11 cases. Rete testis and epididymis cells displayed a heterogeneous co-expression of cytokeratin and vimentin. In double immunohistochemistry, differences in distribution of keratin and vimentin intermediate filaments with predominance of cytokeratins in the apical cytoplasmic regions and of vimentin filaments in the basal portions of the cells were found. Cytokeratin expression preceded the appearance of vimentin: cytokeratin was already detectable in 10-week-old embryos, while weak vimentin immunoreactivity was first seen in 12-week-old embryos and became conspicuous in testes around the perinatal period. In testes of children up to 2 years of age the cytoplasmic distribution of cytokeratin and vimentin was more homogeneous. Predominance of the basal cell portions for vimentin and the apical regions for cytokeratin staining were less pronounced than in adult testes. In the proximal and distal parts of the epididymis a different intermediate filament expression pattern was found with a clear predominance of cytokeratin near the rete.

The angiogenic and fibrogenic activities of fibrin sealant (Tisseel) suggest that it could enhance osteoinductivity during the first days after implantation. To test this hypothesis, the osteoinductivity of sealed and unsealed bone matrix gelatin was tested by implantation into rat abdominal wall muscle pouches for two to ten days. A sponge of collagen with or without fibrin sealant was used for control. Alkaline phosphatase (ALPase) was employed to quantify osteoinduction, and myeloperoxidase (MPO) was used to determine the inflammatory reaction. Plain histologic analysis of bone matrix gelatin implants at Days 2, 4, and 10 did not demonstrate any significant differences between sealed and nonsealed implants. Histologic analysis at Day 21 confirmed the osteoinductivity of the implant. The combination of bone matrix gelatin and fibrin sealant induced an early significant increase in ALPase activity, which could be interpreted as early induction of histologically undetectable osteogenesis. A simultaneous increase in the level of MPO suggested that the bone matrix gelatin/fibrin sealant-induced ALPase reaction might be caused by an inflammatory process, but neutrophil counting did not correlate to the MPO data. Morphometry, however, seems to be insufficient to assay a heterogenous histologic distribution of singular cells. It is proposed that the ALP/MPO ratio may be used as an inflammation-independent marker of osteogenesis. This study does not support the notion of an osteoinductive potency of fibrin sealant per se; however, it is clear that fibrin sealant does not impair bone matrix gelatin-dependent osteoinduction.

The multiple malignancies (MM) autoptically verified at our department within a period of 15 years were reviewed for evidence of a potential iatrogenic induction. Special emphasis was placed on the problems encountered in causally relating metachronic MM's with the treatment of the preceding malignancy and suitable criteria were developed. In 22 (5.6%) of the 392 MM's available for evaluation induction by previous radiotherapy and/or chemotherapy was thought to be likely; that is 40% of all radiological or chemical treated MM. In light of this relatively low number major objections to vitally indicated anticancer treatment appear to be unfounded. The high share of triple malignancies (34.5%) among the MM's presumably induced by treatment constituted a finding of particular interest.

An experimental study in rats was done to investigate the bone-regenerating properties of collagen apatite (Collapat) and to compare it with osteoinduction dependent on osteogenin-containing gelatine (OCG). The test substances were implanted orthotopically (calvarial defect — 7 mm in diameter) and heterotopically (paravertebral muscles, abdominal muscles). The results were evaluated histologically and enzymatically (alkaline phosphatase). Collapat caused neither osteoinduction in the heterotopic site nor healing of the bone defects. Foreign body reaction without new bone formation was encountered. OCG implantation leads to new bone formation in the muscles within 3 weeks, associated with a significant increase in alkaline phosphatase activity, and to extensive new bone formation in the calvarial defect within 4 weeks. The defects did not heal if left empty. The value of clinical application of Collapat appears to be doubtful. Osteoinduction with OCG requires further experimental investigation.

Two methods for the specific visualization of granulocytes (PMNs) in formalin-fixed and paraffin-embedded tissue samples were compared: (1) the well-established enzyme histochemical method using naphthol-AS-D-chloracetate as a substrate; (2) an immunoperoxidase reaction using anti-PMN serum. Both methods are highly selective for visualizing PMNs. The advantages offered by the immunohistochemical method, i.e. better visualization of the remaining tissue structures and potential combination with other staining techniques, are stressed. Problems of PMN visualization in lung tissue and effects of tissue preparation are discussed. For histomorphometric studies the immunohistochemical method is preferable.

The success of a fibrin glue depends on the correct admixture of fibrinogen, thrombin, and the proteinase inhibitor aprotinin. Using a modification of Astrups method we measured the fibrinolytic activity of surgical specimens of human fallopian tube and uterus. Increasing concentrations of aprotinin were added until there was complete inhibition of fibrinolysis. The fallopian tube and the uterus had higher intrinsic fibrinolytic activity than peripheral nerve tissue. At concentrations of between 10 and 20 KIU of aprotinin, fibrinolysis was blocked in all tissues studied.

Multiple organ failure (MOF) was induced by sterile intraperitoneal inoculation of zymosan in the rat. This results in a typical triphasic illness with maximal clinical signs at Days 2 and 14. In this study, granulocyte superoxide production (unstimulated and phorbol myristic acid stimulated) was studied as well as lipid peroxidation (TBAR) in plasma, liver, and lung tissue. Mainly TBAR levels in liver and lung tissue closely correlated with the triphasic clinical illness, while bacteriological data did not. It is concluded that the severe inflammatory response in this experimental model probably is the result of excessive toxic oxygen radical production. The first phase of illness may mainly be due to oxygen radical formation by activated PMN, the third phase of illness to the production of lysosomal enzymes (proteinases) from PMN, and activated macrophages as indicated by elevated N-acetylglucosaminidase levels.

Summary For investigations on the development of regenerative granulation tissue in wound healing a new model using bone spongiosa (“Kieler Spongiosa”) implants is presented. Particular attention was devoted to developing a model which permits studies on wound healing without disturbance by foreign body reaction and infections. Two test groups of rats received four blocks of “Kieler Spongiosa” each in a symmetrical fashion in a paramedian region underneath the dorsal skin. The spongiosa blocks had previously been treated with glutaraldehyde to achieve cross-linkage of the collagenous structures of the surfaces of the spongiosa trabeculae. After one week (group 1) and two weeks (group 2) the animals were sacrificed. The spongiosa blocks were removed, fixed and evaluated in layered serial sections after decalcification. Two blocks which had been removed randomly from the back and front served for morphometric determination of the total volume of bony substance and the developed granulation tissue. Moreover, the cellular composition of the granulation tissue was morphometrically examined with regard to its content of capillaries, granulation tissue cells and inflammatory cells. The two other blocks were examined for DNA and hydroxyproline content of granulation tissue. Comparison of the two experimental groups yielded marked differences in spongiosa space infiltrated by granulation tissue and its composition. Our model was used to assess the influence of systemic administration of adriamycin andlor local application of a fibrin sealant system on granulation tissue formation and its morphologic structure. A total number of 72 rats was divided into four equally sized groups and underwent paravertebral implantation of two blocks of sterile lyophilised “Kieler Spongiosa” for 14 days: (i) In group 0 (F + A) spongiosa blocks filled with fibrin sealant system were implanted and, in addition, the animals underwent systemic administration of adriamycin. (ii) Group 2 (A) underwent systemic administration of adriamycin, without fibrin, on the day of surgery followed by, implantation of the spongiosa blocks. (iii) In Group 1 (F) spongiosa blocks filled with fibrin sealant were implanted without adriamycin administration. (iv) Group 3 (CO) received spongiosa blocks without additives and without adriamycin administration (control group). The results revealed a significant inhibition of wound healing by adriamycin, which could not be prevented by local application of fibrin sealant. In the group treated only locally with fibrin sealant system a significant increase of local granulation tissue cells (mainly fibroblasts) was noted with slight reduction of the amount of granulation tissue and without striking changes in the composition of the inflammatory infiltrate.

Bone matrix gelatin induces bone formation in muscle, and when implanted orthotopically it improves bone repair. Co-60 sterilization of bone gelatin impairs the protein-bound induction mechanisms. Gelatin samples nonirradiated or irradiated by 25 or 50 kGy were implanted into a pouch in the abdominal wall of Sprague-Dawley rats, as well as into a 7-mm calvarial defect. Evaluation was done by histologic studies, histomorphometry of orthotopic implants, and determination of alkaline phosphatase in ectopic implants. Gelatin irradiated with 50 kGy was absorbed in the muscle bed without evidence of any specific host reaction. Irradiation of 25 kGy led to histologically confirmed ectopic bone formation, but the wet weight of the explants was only half that of the nonirradiated control samples. Alkaline phosphatase activity was equal in both of these groups. With orthotopic implantation, neither a histologic nor a morphometric effect was seen with 25 kGy. Loss of osteoinduction with 25-kGy irradiation is apparently masked by osteoconductive mechanisms with orthotopic implantation.

Demineralized bone matrix and bone-matrix gelatin prepared from cortical rat bone, and from cortical and cancellous canine bone were implanted into muscle pouches of nude rats for 6 weeks. Evaluation was done by histology, histomorphometry, and determination of alkaline phosphatase. Rat matrix consistently induced new bone and high phosphatase levels. Canine matrix induced but small amounts of bone and lower phosphatase levels, with cortical matrix somewhat more inductive than cancellous matrix; demineralized cancellous bone matrix from the dog was the only material tested not showing any inductivity. Irrespective of bone type or species, gelatin had clearly higher induction capacity than demineralized bone matrix.

In an extensive series of experimental studies conducted on the common carotid artery in rats, a combined technique involving suture and gluing with human fibrin glue has been developed. In view of the good results achieved, this technique was then applied in 12 patients in whom anastomosis between the superficial temporal artery and a cortical branch of the middle cerebral artery was carried out. This modified surgical technique shortened the duration of operation and also reduced the incidence of complications. Late results, too, seem to be more satisfactory, as far as can be ascertained within the given observation period. 10 patients showed a marked improvement in the clinical neurological findings. Working capacity was completely restored in 5 out of these patients. In one patient, clinical symptoms deteriorated on account of an intra-operative contralateral cerebral haemorrhage. The condition of one patient remained unchanged.

In several reviews (Nadji 1980; Walts and Said 1983; Koss 1984; Chess and Hajdu 1986) the considerable value of the application of immunocytochemical methods in cytology (ICC) has been indicated. However, despite the high number of reports focused on the application of ICC as a diagnostic tool it did not have the enormous practical impact of immunohistochemistry. This may be due to methodological difficulties and the existence of a vast number of different protocols for fixation and storage and the different immunocytochemical techniques used for cytological preparations. Because of inaccuracies in the descriptions of fixation and processing of cell preparations in many papers on ICC, a comparison of data obtained by different authors is impossible or at least questionable.

Immunocytochemistry represents an important tool in diagnostic cytology. Its application, however, is of restricted value due to methodologic difficulties, leading in some cases to false-positive or false-negative results. This study examined the influence of different fixatives and modes of storage of cytologic cell preparations on immunoreactivity using immunoenzymatic techniques. Monoclonal and polyclonal antibodies were used on imprint preparations of breast lesions and tonsils and on bronchoalveolar lavages. A short fixation of the cytologic material with 0.05% glutaraldehyde or a commercially available spray fixative (Merckofix) enabled the immunolocalization of a considerable number of antigens. Optimal demonstration of some antigens, however, required the use of individual fixatives. Drying of cell preparations should be avoided at any step of the procedure to prevent false-positive or false-negative results. Smears stored in methanol or sucrose-MgCl2-glycerol solutions retained their immunoreactivity for several months. The results of this study are compared with those of previous reports, and the reasons for, and means of avoiding, frequent pitfalls in immunocytochemistry are discussed.