Detection of blood Gb3 deposits as a new tool for diagnosis and therapy monitoring in patients with classic Fabry disease.

BACKGROUND: The X-linked Fabry disease (FD) is a multiorgan disorder due to alpha-galactosidase A (α-GAL) deficiency with consequent lysosomal accumulation of globotriaosylceramide (Gb3). We established the immunocytochemical detection of Gb3 in blood cells of FD patients as a new method for FD diagnostics, follow-up and treatment control. METHODS: We enrolled 67 FD patients (37 men, 30 women) and 52 healthy controls (26 men, 26 women). PBMC were isolated from whole venous blood and 3x105 cells were immunoreacted with antibodies against CD77 as a marker for Gb3. Using fluorescence microscopy, the mean percentage of Gb3 positive PBMC was determined by an investigator blinded to subject allocation. As a second method, we qualitatively assessed Gb3 positive cells in blood smears. RESULTS: Gb3 deposits were unequivocally visible in PBMC and in blood smears. Men (P < 0.001) and women (P < 0.01) with classical FD had more Gb3-positive PBMC than healthy controls, whose samples only occasionally showed positive cells. The number of Gb3 positive PBMC was negatively correlated with α-GAL activity and positively correlated with plasma lyso-Gb3 levels. Only the PBMC Gb3 load but not plasma lyso-Gb3 reflected short- and long-term effects of enzyme replacement therapy (P < 0.01). CONCLUSIONS: Gb3 can be visualized in PBMC and blood smears and can be used as a novel marker for diagnostics, follow-up and treatment control in FD.

Hypoxia-stimulated membrane trafficking requires T-plastin.

AIM: Traffic between the plasma membrane and the endomembrane compartments is an essential feature of eukaryotic cells. The secretory pathway sends cargoes from biosynthetic compartments to the plasma membrane. This is counterbalanced by a retrograde endocytic route and is essential for cell homoeostasis. Cells need to adapt rapidly to environmental challenges such as the reduction of pO2 which, however, has not been analysed in relation to membrane trafficking in detail. Therefore, we determined changes in the plasma membrane trafficking in normoxia, hypoxia, and after reoxygenation. METHODS: Membrane trafficking was analysed by using the bulk membrane endocytosis marker FM 1-43, the newly developed membrane probe mCLING, wheat germ agglutinin as well as fluorescently labelled cholera toxin subunit B. Additionally, the uptake of specific membrane proteins was determined. In parallel, a non-biased SILAC screen was performed to analyse the abundance of membrane proteins in normoxia and hypoxia. RESULTS: Membrane trafficking was increased in hypoxia and quickly reversed upon reoxygenation. This effect was independent of the hypoxia-inducible factor (HIF) system. Using SILAC technology, we identified that the actin-bundling protein T-plastin is recruited to the plasma membrane in hypoxia. By the use of T-plastin knockdown cells, we could show that T-plastin mediates the hypoxia-induced membrane trafficking, which was associated with an increased actin density in the cells as determined by electron microscopy. CONCLUSION: Membrane trafficking is highly dynamic upon hypoxia. This phenotype is quickly reversible upon reoxygenation, which suggests that this mechanism participates in the cellular adaptation to hypoxia.

Changes of murine sperm phospholipid composition during epididymal maturation determined by MALDI-TOF mass spectrometry.

After leaving the testis, spermatozoa undergo several important steps of biochemical maturation during the passage through the epididymis, increasing their motility and fertilizing ability. These changes comprise (among others) the modification of the phospholipid composition of the sperm membrane. This process is thought to be important for the achievement of motility and fertilizing capacity. The lipids of the sperm membrane are characterized by a significant content of unsaturated fatty acyl residues, resulting in a high sensitivity against oxidative stress. This is evidenced by the appearance of lysolipids, for example, lysophosphatidylcholine, which acts like a detergent and is normally present in only very small amounts in biological membranes. The epididymis represents a tubular system comprising three main parts (caput, corpus, and cauda), through which the spermatozoa are consecutively transported undergoing distinct maturation stages. Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, we established three striking differences in the lipid composition of murine spermatozoa from the different epididymal regions: in comparison to the caput sperm, sperm from the cauda are characterized by (1) a higher degree of unsaturation (PC 18:0/22:5 and 18:0/22:6 vs. 18:0/20:4 and 18:0/18:1), (2) an enhanced plasmalogen content, and (3) an enhanced content of lysolipids. These changes are likely to be of physiological relevance and potentially useful as diagnostic markers of sperm maturation and acquisition of motility.

[Pancreatic heterotopia as a rare diagnosis of a size increasing gastric tumor]. / Pankreasheterotopie als seltene Differenzialdiagnose eines grössenprogredienten Magentumors.

A 39 years old woman was noted to have a submucosal gastric tumor during gastroscopy for cholecystectomy. She was asymptomatic regarding this tumor. The patient was primarily observed, but the tumor size increased. The lesion was biopsied with a negative result. A local excision with simultaneous endoscopy was done laparoscopically. Malignancy was excluded by immediate sectioning. Histological examination revealed pancreatic heterotopy Type I by Heinrichs- with exocrine and endocrine cells. The patient had an uneventful postoperative course. Eight months after resection the patient was free from recurrence and symptoms.

[The thoracoscopic thymectomy for myasthenia gravis]. / Die thorakoskopische Thymektomie bei der Myasthenia gravis.

UNLABELLED: Myasthenia gravis (MG) is a chronic autoimmune disease that usually responds positively to treatment with thymectomy. Standard surgical procedures have been shown to result in a favorite outcome. The optimal surgical access, however, is under discussion. PATIENTS AND METHOD: In a prospective study conducted between 8/97 and 4/05, 173 patients with MG underwent thoracoscopic thymectomy, for which a left-sided approach was generally applied. An analysis of the intraoperative and postoperative course was performed in 137 patients (8/97 and 12/03), as well as of the impact of the surgical procedure on further development of the disease. The results obtained were compared with those published in the literature, with particular reference to results obtained with open surgery. In 8 patients, the procedure was converted to an anterolateral thoracotomy or sternotomy, so we examined 129 patients, which were operated thoracoscopically. RESULTS: The patients in this study were 93 females and 36 males, with a mean age of 35.8 (range: 9-83) years. The mean preoperative duration of the disease was 22.9 (range: 1-140) months. The duration of operation was 50 to 85 minutes, the blood loss was lower than 100 ml. The mean follow up was 24.7 (range: 1-57) months. Complete remission was noted in 30 patients (23.3 %), and improvements reflected either in a reduction of the required medication, or a decrease in symptom severity, were seen in a total of 86 patients (66.6 %). CONCLUSION: Complete thoracoscopic thymectomy is a technically feasible operation, and as effective as conventional open surgery. Remissions or symptomatic improvements were observed in more than 89 % of the patients. The low morbidity rate, in combination with excellent cosmetic results, has led to increasing acceptance of the operation both by patients and neurologists. Therefore, thoracoscopic thymectomy represents a new, alternative method for treatment of patients with MG.

Invasive staging of non-small cell lung cancer--a prospective study.

OBJECTIVES: Clinical prognosis and treatment schedules of non-small cell lung cancer (NSCLC) are dependent on tumor stage. This explains the importance of an exact pretreatment staging of the primary tumor and lymph nodes especially in locally advanced NSCLC, to differentiate between resectable and non-resectable disease. To assess the lymph node status of the upper mediastinum, the diagnostic value of mediastinoscopy is accepted to be superior to radiological methods. In contrast, thoracoscopy is not yet established as a standard staging tool. PATIENTS AND METHODS: Seventy-three consecutive patients with CT-based suspicion of advanced NSCLC have been investigated as part of a phase II study on neoadjuvant treatment of NSCLC. All patients underwent mediastinoscopy and mediastinal lymph node sampling. In the case of a negative result we performed additional thoracoscopy. RESULTS: In 52.1% (n = 38) of the patients the invasive diagnostic methods led to results that were effectively different from those of the radiological findings. In 11 patients (15.1%) CT-assessed lymph node metastases could invasively not be confirmed, whereas nine patients (12.3%) had positive mediastinal lymph nodes but no corresponding CT signs (diameter <1 cm). The results were achieved by mediastinoscopy in 15 (20.5%) and by thoracoscopy in five (6.8.%) patients. A radiologically unexpected T4 stage has been found in four (5.5%) and a M1 stage in four (5.5%) patients by thoracoscopy. On the contrary, in seven patients a suspected infiltration of mediastinum or parietal pleura could be thoracoscopically excluded. Four patients have been in an unexpected high stage of tumor progression at the moment of diagnostic procedures and therefore have been included in palliative therapy schedules. Ten patients have been 'overstaged' by radiological methods and benefited from a primarily curative resection after invasive staging. CONCLUSIONS: Of the 73 prospectively studied patients with locally advanced NSCLC, 12 (16.4%) have been staged too low and 13 (17.8%) too high. If exclusively staged by radiological methods, about 34% of lung cancers have been classified incorrectly. Therefore, these tools are not a sufficient basis for diagnosis of stage III NSCLC disease. Mediastinoscopy with consecutive thoracoscopy is an essential part of the therapeutic planning in locally advanced NSCLC, and results are significantly superior to clinical staging.

Effects of supplementation with high linoleic or oleic cracked safflower seeds on postpartum reproduction and calf performance of primiparous beef heifers.

Primiparous Angus x Gelbvieh (n = 36) rotationally crossed beef cows (initial BW = 487.9 +/- 10.5 kg, body condition score = 5.5 +/- 0.02) were utilized to determine effects of supplemental safflower seeds high in linoleic (76% 18:2) or oleic (72% 18:1) acid on cow BW change, body condition score, milk production and composition, calf weight gain, cow serum metabolites, and metabolic hormones. On d 3 postpartum, cows were randomly assigned to one of three isonitrogenous dietary supplements with equal total quantity of TDN: corn-soybean control supplement (n = 12); high-linoleate safflower seeds (n = 12); or high-oleate safflower seeds (n = 12). Safflower-seed supplements were formulated to provide 5% DMI as fat. Supplements were individually fed from d 3 postpartum through 90 d postpartum. Cows had ad libitum access to native grass hay (7.8% CP), trace-mineralized salt, and water. Date of parturition was evenly distributed across treatments with all cows calving within 14 +/- 0.8 d. There were no differences (P = 0.65) in total OM intake among treatments. Although cow BW change did not differ (P = 0.33) by treatment, supplementation influenced cow body condition score (P = 0.02) with linoleate-supple-mented cows in higher (P = 0.005) condition overall than oleate-supplemented cows (5.1 +/- 0.06 vs 4.9 +/- 0.06). Twenty-four-hour milk production did not differ (P = 0.68) among treatments. Percentage milk fat was not different at d 30; however, at d 60 and d 90 percentage milk fat was greater (P ( 0.05) in control and oleate-supplemented cows than in linoleate-supplemented cows. Calf BW gains (P = 0.27) and adjusted 205-d weights (P = 0.48) were not affected by supplement treatment. Supplementation did not influence serum concentrations of glucose (P = 0.38), NEFA (P = 0.61), GH (P = 0.29), IGF-I (P = 0.81), insulin (P = 0.26), or IGF-I binding proteins (P > or = 0.11). Days to conception did not differ (P = 0.40) among treatments. Although overall productivity of the primiparous cows and their calves was not altered by safflower-seed supplementation, differential effects were noted between supplements. Oleate supplementation increased percentage milk fat at d 60, and cow body condition score was lower than in linoleate-supplemented cows. Linoleate-supplemented cows had greater body condition scores by 90 d postpartum than either corn-soybean- or oleatesupplemented cows.

[Minimally invasive concepts in esophageal surgery--thoracoscopic anastomosis. An animal experiment study]. / Minimal-invasive Konzepte in der Osophaguschirurgie--thorakoskopische Anastomosierung. Eine tierexperimentelle Studie.

Based on animal trials the presented study describes two versions of thoracoscopic oesophageal anastomosis within the scope of abdomino-thoracoscopic oesophagectomy. This experimental approach is considered to provide a solution for the oncological problem to salvage the tumor bearing oesophagus. We describe the procedure of intrathoracic stapler anastomosis under thoracoscopic vision. By using a laparoscopic purse string suture clamp we were enabled to prepare the proximal oesophageal stump for anastomosis. A circular stapling instrument turned out to be very suitable for the thoracoscopic use. The final evaluation of the importance of minimally invasive surgery for the resection of oesophageal carcinomas should not be given until multicenter studies are performed.

Hardware-based rendering of full-parallax synthetic holograms.

We present a method for efficiently calculating the interference of complex-valued two-dimensional wave patterns that is useful during the generation of synthetic holograms. These patterns are represented as a special kind of images (textures), and the interference is calculated in a computer graphics rendering process. This enables us to leverage hardware support for holographic imaging that is implemented in many state-of-the-art computer workstations. Using this approach, we gain a speedup of a factor of 60-90 compared with conventional calculation methods for interfering wave patterns. Our method is evaluated numerically, examples are shown, and the program code is outlined.

MRI, CT, US and ERCP in the evaluation of bile duct hamartomas (von Meyenburg complex): a case report.

A case of multiple bile duct hamartomas (von Meyenburg complex) of the liver accompanied by exudative and in part necrotizing pancreatitis is presented. Magnetic resonance imaging (fat suppressed, T2-weighted images with prolonged echo time) could exclude diffuse tumor infiltration of the liver, which had not been possible with CT, sonography, or ERCP. To our knowledge, no comparable case has been reported.

[Surgical therapy of chest wall infiltrating local recurrence of breast carcinoma]. / Zur chirurgischen Therapie des brustwandinfiltrierenden Mammakarzinom-Lokalrezidivs.

Chest wall resection and--reconstruction for malignant tumors have to fulfill oncological, functional and cosmetic demands. We report on 19 operated patients with chest wall recurrence or metastasis from breast cancer. Resection included excision of the thoracic shield (n = 14), additional partial resection of breast bone (n = 5) and of clavicle (n = 1). Chest wall stability was achieved by PTFE-patches and lyophilized bone, soft tissue reconstruction was performed by myocutaneous flaps. Resection rate was 95%, morbidity rate was 23%, we saw no in-hospital deaths. 1-year-survival was 95%.

[Determinants of long-term prognosis following prosthetic mitral valve replacement]. / Determinanten der Langzeitprognose nach prothetischem Mitralklappenersatz.

UNLABELLED: Data of 235 patients with chronic mitral valve disease (NYHA class III or IV) were analyzed retrospectively. Mitral valve replacement was performed in all patients between 1974 and 1983, 46 patients underwent additional tricuspid valve repair during the same operation. Preoperatively, right and left heart catheterization as well as coronary angiography were carried out in all patients. Operative mortality was 8%. For all patients 5 year survival rate was 76% and 10 year survival rate was 72%. Patients with pure mitral insufficiency had a poorer long-term prognosis than patients with mitral stenosis or patients with mixed mitral valve disease (5 year survival rate 54 versus 79%, p less than 0.0014). Patients with previous closed mitral commissurotomy had a poorer long-term prognosis than patients without previous surgery (5 year survival rate 62 versus 81%, p less than 0.0019). Age, NYHA class, associated tricuspid valve repair, pulmonary vascular resistance and mean right atrial pressure had no effect on survival. CONCLUSION: Long-term survival is generally good after mitral valve replacement, but pure mitral insufficiency and previous closed mitral commissurotomy are risk factors for long-term prognosis.

[Surgical indications in mitral valve stenosis, combined mitral valve defect or mitral valve insufficiency. Long-term prognosis of operated or drug-treated patients]. / Zur Operationsindikation bei Mitralklappenstenose, kombiniertem Mitralklappenfehler oder Mitralklappeninsuffizienz. Langzeitprognose operierter oder medikamentös behandelter Patienten.

Data obtained from 683 patients with mitral valve disease, NYHA-class III or IV, were retrospectively studied by means of a multivariate Cox regression analysis. Based on symptoms and hemodynamic findings, surgical intervention had been recommended for all patients: closed mitral commissurotomy (n = 361), prosthetic mitral valve replacement (n = 241) and prosthetic mitral valve replacement together with a corrective procedure for the tricuspid valve (n = 81). While the majority of patients underwent surgery during the observation period (n = 528), a substantial number of patients continued on medical treatment (n = 155). The mean observation periods were 52, 49 and 31 months, respectively, in the three collectives. Surgically treated patients in whom closed mitral commissurotomy had been recommended had a better prognosis (p less than 0.0003) than those treated medically (five-year survival rate 89% vs. 63%). Age, clinical severity, previous mitral commissurotomy, pulmonary vascular resistance and right atrial mean pressure had no significant influence on prognosis. In patients in whom prosthetic mitral valve replacement had been recommended, surgical treatment led only to tendencial improvement in prognosis as compared with those treated medically (five-year survival rate 78% vs. 61%). Factors with an unfavorable influence on prognosis were age more than 49 years (p less than 0.05), pure mitral regurgitation (p less than 0.001), NYHA-class IV (p less than 0.02) and right atrial mean pressure in excess of 4 mm Hg (p less than 0.01). In patients in whom prosthetic mitral valve replacement together with a corrective procedure for the tricuspid valve had been considered necessary, surgical treatment had no significant influence on prognosis as compared with those treated medically (five-year survival rate 57% vs. 53%). Patients in whom previous mitral commissurotomy had been performed had an extremely poor prognosis (p less than 0.001). Pulmonary vascular resistance was significantly reduced both after mitral commissurotomy as well as after prosthetic mitral valve replacement; this was associated with a significant decrease in right atrial mean pressure and increase in right ventricular ejection fraction. The indication for closed mitral commissurotomy, thus, appears established in patients with symptoms of class III or IV clinical severity. The indication can be established generously since the surgical mortality is low and long-term prognosis is good.(ABSTRACT TRUNCATED AT 400 WORDS)