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α-Gal syndrome results from sensitization to the carbohydrate epitope galactose-α-1,3-galactose (α‑gal). The allergen occurs in mammalian meat and innards, but also in other foods and medical products of animal origin. Allergic reactions generally occur delayed after allergen intake with a latency period, depending on the individual tolerance threshold and the influence of cofactors. Details in the patient's medical history can help to establish the suspected diagnosis of α‑gal syndrome. Confirmation of the diagnosis requires the expertise of specialists, experienced with the implementation and interpretation of in vitro and in vivo diagnostic tests. Whereas skin prick testing with commercial whole-meat extracts often does not provide reliable results, allergen-specific IgE (α-gal) is generally detectable in affected patients. Cell-based tests such as the basophil activation test are currently only employed in an experimental setting. To evaluate, whether a sensitization is clinically relevant, an in-patient oral food challenge should be performed, using for example cooked pork or porcine kidney in addition to suspected cofactors.

Fungi of the genus Aspergillus are ubiquitously present. Even though humans inhale Aspergillus spores daily under natural conditions, Aspergillus-associated pulmonary diseases only occur under special circumstances. Whether an Aspergillus-associated disease develops and which type of Aspergillus-associated disease develops depends on the constitution of the host. The spectrum of Aspergillus-associated pulmonary diseases ranges from allergic diseases, such as hypersensitivity pneumonitis to allergic infectious diseases, such as allergic bronchopulmonary aspergillosis (ABPA) and bronchocentric granulomatosis (BG) to infectious diseases, such as invasive (IA) or semi-invasive aspergillosis (SIA) and chronic pulmonary aspergillosis (CPA). Identification of Aspergillus spp. from sputum or bronchopulmonary secretions is not sufficient for a definitive diagnosis of Aspergillus-associated infections. The gold standard is the identification of Aspergillus spp. from lung tissue by culture or by histopathological methods; however, in clinical practice the decision to initiate antifungal therapy is more often based on immunological methods, such as the detection of Aspergillus-specific IgG antibodies from peripheral blood or galactomannan antigens from bronchoalveolar lavages. Acute IA or SIA infections have a high mortality and require immediate antifungal therapy. With rare exceptions CPA cannot be cured by medicinal therapy alone; however, active CPA can be brought into remission with antifungal therapy. Eradication of Aspergillus in CPA can as a rule only be successful using a combined antimycotic and surgical intervention.

Genomanalysen deuten darauf hin, dass in dem Haloarchaeon Haloferax volcanii Galactose über einen DeLey-Doudoroff- (DD) Weg abgebaut wird. Der Transport von Galactose und die beteiligten Enzyme des Galactose-Abbaus wurden bisher nicht untersucht. Genomanalysen des Haloarchaeons Halorhabdus utahensis deuten darauf hin, dass Galactose, anders als für H. volcanii postuliert, über einen Leloir-Weg abgebaut wird. Untersuchungen zum Galactose-Abbau in H. utahensis wurden bisher nicht durchgeführt. Weiterhin deuten Untersuchungen darauf hin, dass Glucose in H. volcanii über einen semiphosphorylierten Entner-Doudoroff- (spED) Weg abgebaut wird. Das Schlüsselenzym dieses Wegs, die KDPG-Aldolase, wurde bisher nicht detailliert charakterisiert. Die Enzyme der Oxidation von Glycerinaldehyd-3-Phosphat (GAP), einem Intermediat des Galactose- und Glucose-Abbaus in H. volcanii wurden nicht untersucht. In der vorliegenden Arbeit wurden der Transport und der Abbau von Galactose, sowie die transkriptionelle Regulation der Gene des Galactose-Abbaus in H. volcanii untersucht. In H. utahensis wurde das Schlüsselenzym des Leloir-Wegs charakterisiert. Außerdem wurde die KDPG-Aldolase, sowie die Enzyme der GAP-Oxidation zu 3-Phosphoglycerat in H. volcanii detailliert untersucht. Im Folgenden werden die in der vorliegenden Arbeit erzielten Ergebnisse zusammengefasst... Genome analyses suggest that in the halophilic archaeon Haloferax volcanii galactose is degraded via a DeLey-Doudoroff pathway (DD). The transport of galactose and the enzymes involved in galactose degradation in H. volcanii have not been characterized so far. Genome analyses of the halophilic archaeon Halorhabdus utahensis suggest, that galactose, in contrast to H. volcanii, is degraded via the classical Leloir-pathway. Enzymes involved in galactose degradation in H. utahensis have not been analyzed so far. Furthermore, recent studies indicate that glucose is degraded via a semiphosphorylative Entner-Doudoroff pathway (spED). The key enzyme of this pathway, KDPG aldolase, has not been characterized in detail. Also, the enzymes involved in oxidation of glyceraldehyde-3-phosphate (GAP), an intermediate of galactose and glucose catabolism in H. volcanii, have not been analyzed so far. In this thesis the transport and degradation of galactose as well as transcriptional regulation of the genes involved in galactose degradation in H. volcanii were investigated. In H. utahensis the key enzyme of Leloir pathway was characterized. Moreover, KDPG aldolase, the key enzyme of glucose degradation as well as enzymes catalyzing GAP oxidation to 3-phosphoglycerate were investigated in detail in H. volcanii. The following results were obtained in this thesis...

Diseases caused by Aspergillus spp. are difficult to diagnose and thus require supplementary serological assays. This is the result of a selective review of the relevant literature with special regard to recent guidelines. In addition to conventional diagnostic tools (radiology, microscopy, culture) the measurement of the following serological markers is recommended, depending on the clinical type of aspergillosis: Invasive and chronic necrotising aspergillosis: Aspergillus-galactomannan antigen. Test format: EIA using the rat MAb EB-A2. Cut-off 0.5 (index). Monitoring of high risk patients: Twice weekly. Aspergillus-IgG (test format EIA) as confirmatory assay after recovery of the leukocyte function under therapy. Aspergilloma: Aspergillus IgG. Test format: EIA. Allergical aspergillosis: Aspergillus IgE. Test format: RAST. Galactomannan antigen detection rates high in the diagnosis of invasive aspergillosis. The evaluation of Aspergillus nucleic acid amplification assays is pending.

Fungal infections are of great relevance in surgical intensive care and Candida species represent the predominant part of fungal pathogens. Invasive aspergillosis is also relevant especially in patients with chronic pulmonary diseases. It is crucial for therapy success to begin adequate antifungal treatment at an early stage of the disease. Risk stratification of individual patient symptoms is essential for therapy timing. In case of suspected or proven candida infection, fluconazole is the agent of choice when the patient is clinically stable and no azoles have been administrated in advance and the local epidemiology makes azol resistance unlikely. For clinically instable patients with organ dysfunction the echinocandins serve as primary therapy because of their broad spectrum and reasonable safety profile. Due to a relevant proportion of azole resistant Candida species, susceptibility testing should be done routinely. Depending on the species detected de-escalating to an azole is feasible if organ dysfunctions have resolved. An invasive aspergillosis is primarily treated with voriconazole.

In this work, the binding characteristics of a Tn and a TF-Antibody and two lectins, HAA from Helix aspersa and Jacalin towards IgA1 O-glycans were characterized in detail. The aim was to evaluate existing techniques for the detection of undergalactosylated IgA O-glycans in IgA nephropathy and to develop and analyse new detection methods. A set of serum samples from IgAN patients and controls was analyzed with the antibodies and the HAA lectin. However, no significant difference in the glycosylation profile could be found. A new technique for the detection of undergalactosylated IgA O-glycans was developed, that employs the enzymatic labeling of undergalactosylated glycans. Therefore, an enzyme that was able to transfer galactose onto free GalNAc sites on IgA1 had to be cloned. For this purpose, three homologous core 1 beta1,3 galactosyltransferases, that transfer galactose onto GalNAc from human, drosophila melanogaster and E. coli were recombinantly expressed and characterized in detail. The human enzyme was only formed in inclusion bodies and showed almost no activity. The bacterial enzyme was also formed in inclusion bodies. The solubility and activity of this enzyme could be greatly enhanced by expressing it as fusion protein with a propeptide from a lipase of staphylococcus hyicus. However, it was not able to transfer galactose onto IgA1 O-glycans. The enzyme from drosophila melanogaster showed a high activity when expressed in Hi5 insect cells and was the only one able to transfer galactose onto IgA1 O-glycans. In the first approach, unlabeled galactose was transferred by the drosophila C1 GalT onto free GalNAc moieties on IgA1. The binding profile of the Tn- and TF-antibody and the HAA lectin to IgA O-glycans was analyzed before and after the transfer of galactose. This should establish a homogenously galactosylated glycan standard. The comparison of the galactosylation profile before and after the transfer should show a great difference in undergalactosylated samples and a little difference in normally galactosylated samples and thus identify IgAN patients. Unfortunately, this method, also was not able to show a significant difference in the IgA glycosylation of IgAN patients and controls. This was greatly due to the poor specificity of the antibodies and lectin for their sugars. Secondly, free GalNAc moieties on IgA1 from the set of serum samples from IgAN patients and controls were labeled by the drosophila C1GalT with radioactive galactose (3H galactose). By this technique, no significant difference in the glycosylation pattern of IgAN patients and controls could be demonstrated. It seemed that enzyme kinetics prevented a complete labeling of all free GalNAc moieties. The aim to develop and evaluate different methods for the analysis of IgA1 O-glycans was achieved. However, none of the analysed methods was able to discriminate between IgAN patients and controls significantly. No method that was suitable as a sero-diagnostic test for IgAN could be developed.

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Bei dem Contrast Burst Depletion Imaging (CODIM) wird die Gewebsperfusion nach dem Contrast Burst Imaging Modus über eine Serie kontrastmitteldestruierender Ultraschallimpulse gemessen: Das im Intervall zwischen zwei Insonationen in das Messvolumen einströmende Kontrastmittel wird destruiert und über die dabei frei werdende Schallenergie gemessen. Die Modellfunktion des Verfahrens beschreibt einen exponentiellen Abfall der zu den Insonationszeitpunkten gemessenen Werte. In der vorliegenden Arbeit wurden 15 gefäßgesunde Probanden nach der CODIM-Methode mit dem Echokontrastmittel Levovist® untersucht. Es konnte gezeigt werden, dass der aus der Modellfunktion errechnete Perfusionskoeffizient ein von der jeweiligen Untersuchungstiefe unabhängiges, semiquantitatives Maß der Gewebsperfusion darstellt.

Bei dem Contrast Burst Depletion Imaging (CODIM) wird die Gewebsperfusion nach dem Contrast Burst Imaging Modus über eine Serie kontrastmitteldestruierender Ultraschallimpulse gemessen: Das im Intervall zwischen zwei Insonationen in das Messvolumen einströmende Kontrastmittel wird destruiert und über die dabei frei werdende Schallenergie gemessen. Die Modellfunktion des Verfahrens beschreibt einen exponentiellen Abfall der zu den Insonationszeitpunkten gemessenen Werte. In der vorliegenden Arbeit wurden 15 gefäßgesunde Probanden nach der CODIM-Methode mit dem Echokontrastmittel Levovist® untersucht. Es konnte gezeigt werden, dass der aus der Modellfunktion errechnete Perfusionskoeffizient ein von der jeweiligen Untersuchungstiefe unabhängiges, semiquantitatives Maß der Gewebsperfusion darstellt.

Galaktose-Gegentransportmessungen an Plasmamembranvesikeln präpariert aus mit Galaktose induzierten Zellen der Nullmutante RE700 ergaben folgende Km-Werte: 6,4, 6,5 und 5,3 mM. Damit ist zum erstenmal eindeutig der Mechanismus des ?facilitated diffusion? Transportes und die Kinetik des Galaktosetransporters Gal2 der Hefe bestimmt worden. Der Galaktosegegentransport an Plasmamembranvesikeln präpariert aus der Nullmutante RE700 nach Galaktose-Induktion zeigt im Vergleich zu dem mit Glukose ein deutlich niedrigeres Gegentransportmaximum, was bereits auf eine geringere Affinität von Galaktose zum Gal2 hinweist. Die entsprechenden Km-Werte aus sechs Versuchen des Glukose-Gegentransportes waren 3,9, 2,0, 4,0, 2,0, 2,2 und 2,3 mM. Der hieraus berechnete Mittelwert von 2,7 ± 0,4 mM entsprach damit der Hälfte des Km-Wertes aus dem Galaktosegegentransport mit 6,0 mM. Eine ebenso große Differenz der Km-Werte zwischen dem Transport von Galaktose und Glukose konnte bei initialen Aufnahmeversuchen an Zellen der Nullmutante RE700 nach Galaktose-Induktion erhoben werden. Aus 28 Einzelversuchen mit Galaktose als Transportsubstrat ergab sich ein Km-Wert von 3,7 ± 0,3 mM, der damit doppelt so hoch wie der bei der initialen Glukoseaufnahme lag mit einem Km-Wert von 1,8 ± 0,1 mM (46 Versuche). Im t-Test (?two-tailed?) erwies sich dieser Unterschied als hochsignifikant mit einem P-Wert < 0,0001. Der niedrige Km-Wert von 1,8 ± 0,1 mM bei der initialen Glukoseaufnahme bestätigt den berechneten Km-Wert aus den Gegentransport- versuchen von Maier mit 1,5 mM und liegt auch mit dem hier erhobenen Km-Wert von 2,7 ± 0,4 mM im Einklang. Der Km-Wert des initialen Galaktosetransportes beim Wildtyp 996A nach Galaktose-Induktion betrug in 8 Einzelversuchen 4,3 ± 0,7 mM und war somit nicht signifikant verschieden von dem entsprechenden Km-Wert der Nullmutante RE700 mit 3,7 ± 0,3 mM. Die niedrigen Km-Werte für Glukose mit 1,8 mM am Gal2-Transporter der Hefe sind in Übereinstimmung mit entsprechenden Km-Werten des menschlichen Glukosetransporters Glut1 aus der Literatur mit 1,6 und 2,4 mM [Stein90] [Fuhrmann89(1)]. Dagegen sind die Km-Werte für den Galaktosetransport mit 6 und 3,7 mM am Gal2 Transporter der Hefe deutlich verschieden gegenüber dem Km-Wert von 12 mM beim menschlichen Glut1 Transporter. Ein weiterer Unterschied besteht darin, dass der Hefetransporter Gal2 kein asymmetrisches Transportverhalten wie der menschliche Glut1 beim Erythrozyten aufweist. Damit ist nicht nur die Struktur sondern auch die Funktion beider Transporter verschieden. Durch die Anwesenheit von Glukose (?Glukoseshift?) wird eine Repression von Enzymen hervorgerufen. Beim Wildtyp 996A verschwindet der induzierte Galaktosetransporter Gal2 wieder vollständig. Der Abbau erfolgt exponentiell (Korrelationskoeffizient 0,955) mit einem K-Wert von 0,36 ± 0,06 und einer Halbwertzeit von 1,92 Stunden, sodass nach etwa 8 Stunden fast kein Galaktosetransport mehr nachzuweisen ist. Dieses Verhalten und der zeitliche Ablauf entsprechen in etwa den Angaben aus der Literatur [Galindez83] [Horak97]. Im Vergleich zu den Wildtyp-Hefezellen zeigt die Nullmutante RE700 beim ?Glukoseshift? ein signifikant anderes Verhalten, es wird nämlich nur etwa die Hälfte der induzierten Galaktosetransporter abgebaut. Dabei strebt die Abnahme der Galaktosetransporter auf ein Plateau zu. Es werden in diesen Versuchen 40 bis 53% der Galaktosetransporter nicht abgebaut und bleiben aktiv. Dabei ist die Geschwindigkeit des Teilabbaues der Galaktosetransporter schneller als beim Wildtyp 996A. Auf Grund der Streuung lässt sich jedoch hierbei keine exakte Kinetik bestimmen. Das Phänomen der Plateaubildung wurde in dieser Weise bisher noch nicht beobachtet und es ist nicht bekannt, warum die Nullmutante RE700 ihre Galaktosetransporter nicht vollständig abbaut wie Wildtypen der Hefen. Möglicherweise ist durch das Entfernen der Gene für die Glukosetransporter das empfindliche Zusammenspiel der verschiedenen Leloir-Enzyme bei der ?Downregulierung? des Gal2 in der Zelle gestört. Es ist außerdem vorstellbar, dass durch Ausfall der Glukosetransporter als Regulativ bei der Nullmutante ein noch nicht bekannter ?Selbsterhaltungstrieb? die Katabolitinaktivierung des Galaktosetransporters nach gewisser Zeit stoppt. Zur Abklärung dieses interessanten Phänomens sind weitere Versuche notwendig., Regulation and Kinetic Characterization of the Galactose Transport in HXT mutant RE700 Saccharomyces cerevisiae.

Die Arbeit befasst sich mit Methoden der Erfassung metabolischer Funktion einer dynamischen Zellkultur in Bioreaktoren zur klinischen Verwendung als Leberunterstützungssystem. Der Fokus liegt auf der Etablierung von Tests zur Qualitätssicherung als effiziente In- Prozess Kontrollen während der Bereitstellungsphase der Zellkulturen. Weitere Ziele sind ein Beitrag zu Wirksamkeitsnachweis und zur Charakterisierung des Systems im Rahmen von Zulassungen als "Biologic". Bei der experimentellen Untersuchung unter Verwendung von Leberzellkulturen aus porcinen Organen wurden der Lidocain- MEGX Test, die Elimination von Galaktose und Sorbitol sowie die Elimination von Ammoniak und die Synthese von Harnstoff, Lactat und Albumin berücksichtigt. Dies erfolgte mittels Bolus- und Fliessgleichgewichtsuntersuchungen sowie Konzentrationsbestimmungen. Die Ergebnisse zeigten für alle Parameter einen hochsignifikanten Unterschied zu Zellfreien Bioreaktoren, sowie einen typischen Verlauf, der in eine initiale Adhäsionsphase, eine stabile Kulturphase und eine darauf folgende erweiterte Phase mit langsamer Abschwächung der Zellleistungen gegliedert werden kann. Die Parameter erwiesen sich prinzipiell alle geeignet für den Einsatz in der Qualitätssicherung der dynamischen Zellkultur. Ein kombiniertes Untersuchungsschema bestehend aus Parametern die die Integrität der Zellmembranen reflektieren, wie die Freisetzung von Enzymen, sowie metabolischen Parametern wie Lidocain, Galaktose, Sorbitol als auch Syntheseparameter, für Proteine Albumin, als Hepatozyten typische Leistungen die Synthese von Harnstoff und der Abbau von Lactat ist in der Lage einen ausreichend umfassenden Einblick in den Status der dynamischen Zellkultur zu liefern. Objectives of this work were to find easy to handle every day quality assessment procedure for bioreactors that are intended for clinical trials, further more usefulness of these parameters in characterization and standardization of the Liver Support System. Parameters for evaluation had been: Lidocaine, Megx, Sorbitol, Galactose, Urea, Albumin and Lactate. Bolus and Continuous metabolic liver functions tests had been done, for synthesis and detoxification parameters liberation was measured by concentration. Highly significant difference between bioreactor with primary porcine liver cells and cell-free devices was shown by t-tests. Differences to other groups like infected bioreactors had been demonstrated with ANOVA. Results show typical course over the culture period that can be categorized into a initial phase of cell reorganization, a stable culture phase and an extended phase with slow decay. All parameter proved to be suitable for daily routine quality assessment in dynamic cell culture systems. The combination of parameters reflecting different specific cellular function is able to give more comprehensive insights in the status of the cell culture.

\(\textbf {Problem:}\) Seit 1997 wurde der Nutzen von Kontrastmitteln (KM) in Verbindung mit Harmonic Imaging zur Darstellung der Gehirnperfusion untersucht. Ziel dieser Studie war es, zwei neue destruktive Methoden (Contrast Burst Imaging (CBI) und Time Variance Imaging (TVI)) im Vergleich zu Contrast Harmonic Imaging (CHI) zu evaluieren. \(\textbf {Methode:}\) Die Untersuchungen erfolgten an 9 gesunden Probanden in nach i.v. Bolusgabe von 4g Levovist®. Anhand von Zeit-Intensitätskurven konnten Maximalintensität und Zeit bis zum Erreichen dieser für einzelne Gehirnregionen in einer axialen diezephalen Schicht ermittelt werden. \(\textbf {Ergebnis:}\) Alle Methoden zeigten gute Darstellungen von KM in der Mehrheit der untersuchten Regionen (ROI). Dabei wies CBI die besten Ergebnisse auf, gefolgt von TVI und CHI. \(\textbf {Diskussion:}\) Die Studie zeigt zum ersten Mal, dass CBI und TVI die Diagnostik parenchymatöser cerebraler KM-anreicherung ermöglichen, welche die diagnostische Sensitivität von CHI übersteigt.

Increased plasma galactitol levels may lead to development of bilateral pediatric cataract.A 3-year-old boy was found to suffer from a bilateral zonular cataract. Extracapsular lensectomy with posterior capsulotomy, transpupillar anterior vitrectomy and posterior chamber lens implantation were performed during a 4-month-interval.The epimerase-activity in red cells of the index patient was found to be significantly decreased (11.2 mumol/h/g Hb; normal range; 19-35). From other family members, such as the brother (16.8), the father (16.0) and the grandfather (15.6), a diminished red cell activity was observed. The mother whose epimerase activity was considerably lower than that of the above mentioned family members (13.3) showed also a zonular bilateral cataract.Investigation of enzymes and polyols of galactose metabolism as well as consultation of the concerned families are recommended for clarification of cataract development.

A new sandwich antigen enzyme-linked immunosorbent assay (ELISA) for the detection of Aspergillus galactomannan in blood plasma was compared with the latex-agglutination test by investigation of six patients with histologically proven invasive aspergillosis. The ELISA gave positive results in all six patients while the latex test recognized 5/6 as positive. On an average the antigen ELISA gave positive results 19 days earlier than the latex test.

Prospective comparison of B-mode-, plain and signal-enhanced colour Doppler ultrasound to evaluate new criteria for assessment of the tumour status of enlarged cervical lymph nodes.Colour Doppler sonography was performed before and after application of d-galactose in 35 patients with suspected lymph node metastases of the neck. Sonomorphology was analysed qualitatively, maximal flow rate, pulsatility index and resistance index were determined quantitatively.Signal enhanced Doppler revealed a typical vessel morphology of the lymph nodes facilitating differential diagnosis. In 23%, vascularisation of the lymph nodes was only visible after application of the signal-enhancing agent (benign: 44%, malignant: 5%). The measurement of flow velocity and the calculation of RI and PI did not lead to a reliable differentiation between malignant and benign lymph nodes whereas B-mode sonography yielded highly valid results.The application of d-galactose facilitates differentiation between malignant and benign lymph nodes.

Viral RNA is detectable in the serum of the majority but not all patients with chronic hepatitis C. Whether the viremic form differs from the non-viremic form of the disease is unknown. We therefore compared histology (modified Knodell score) and liver function (conventional liver function tests, galactose elimination capacity and aminopyrin breath test) of viremic (n = 45) and non-viremic (n = 37) patients with chronic hepatitis C. Neither the total histologic score, nor any of the individual histologic parameters assessed differed significantly in serum HCV RNA positive and negative patients. Compared to non-viremic subjects, patients with detectable HCV RNA in serum had slightly higher transaminases (p = ns), lower serum albumin (p0.05) and decreased galactose elimination capacity (p0.05). This trend towards more severe functional hepatic impairment in serum HCV RNA positive patients persisted when cirrhotics and non-cirrhotics were analyzed separately; it fell just short of reaching statistical significance, however, most probably due to the small number of subjects per patient group. The median age of serum HCV RNA positive cirrhotics was 17 years, and that of serum HCV RNA negative cirrhotics 22 years higher than that of the respective non-cirrhotics. Biochemical (transaminases) and histological (intralobular and piece-meal necrosis) markers of disease activity, as well as metabolic functional reserve (aminopyrin breath test, galactose elimination capacity) and histologic severity of fibrosis correlated only loosely (Rs = 0.24-0.56), albeit significantly (p0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Doppler blood flow measurements allow differentiation of malignant lesions, that are characterised by an abnormal vascularity. The accuracy of this method is limited due to the physiological flow variation in normal breast parenchyma and in tumours. Magnetic resonance imaging in combination with contrast agents can also be used for flow assessment by measurement of the dynamic contrast enhancement. Recently, a new echo contrast agent (Levovist) has been developed, which survives pulmonary transit and increases the Doppler signal intensity after intravenous injection. Fourteen malignancies with low vascularity were selected for the study. Twenty-two seconds after injection of contrast agent, the Doppler signal intensity increased dramatically. This demonstrates, that echo contrast agents might improve the diagnosis of breast lesions in case of non-conclusive Doppler findings.

The prognostic value of different conventional liver tests was compared with that of the galactose elimination capacity (GEC) in 37 patients with chronic active hepatitis. During a mean observation period of 4.5 years, 12 patients died of the disease. None of the conventional liver tests (transaminases, prothrombin time, albumin, Child-Pugh classification) discriminated between survivors and nonsurvivors. In contrast, GEC discriminated between survivors and nonsurvivors (p less than 0.04). The best prognostic criterion was a decrease in GEC, since only patients with a decrease died of the disease. The positive and negative predictive values of this criterion were 100 and 60% respectively. This concept could prove useful for stratification purposes in therapeutic trials, as well as in assessment of novel forms of treatment of chronic active hepatitis.

The measurements were accomplished by using an experimental setup in which an intestinal loop with adjacent mesentery of an anaesthetized rat was immersed in a buffer-containing cuvette. After i.v. injection of Evans' Blue (EB) the extravasation of the dye, which is bound to plasma albumin, was measured colorimetrically. From the curves recorded it could be derived that after i.v. injection of 2 or 4 g or infusion of 6 g/h glucose (Gk) per kg body weight the permeation of albumin-bound EB was increased by a factor of about 1.9, 3.1 and 2.0 respectively. As mechanism of these effects it is assumed that Gk (but also galactose and saccharose in control experiments) act by osmotically shrinking the endothelial cells and opening their junctions.

The capsule material of Staphylococcus aureus strain 1193/74 could be separated by precipitation with trichloroacetic acid and ethanol as well as by chromatography on DEAE-cellulose into 13 fractions. All fractions contained saccharides and uronic acids as well as amino acids and appeared in their qualitative composition rather similar. However, in quantitative composition and in chromatographic behaviour a rather high degree of heterogeneity could be observed. No clear cut separation of protein and polysaccharide material could be achieved in any fraction. It is supposed, therefore, that the capsule material does not represent merely an acidic polysaccharide, but contains certain amounts of amino acids or peptides varying in a rather wide range.

1. Starting from the spectrophotometric method of Ballard optimal reaction conditions for measurements of galactokinase in piglet liver were systematically studied. These are (final conc. in the test): 100 mM triethanolamine-HCl buffer, 33 mM KCl, 16.5 mM NaF (inhibiting ATPase), 5 mM cysteine hydrochloride, 0.33 mM NADH2, 1 U pyruvate kinase and lactic dehydrogenase, 0.5 mM phosphoenolpyruvate, 1.5 mM galactose, 0.5 mM ATP and 1 mM MgCl2, final pH 7.5. 2. An optimal substrate concentration, a Mg: ATP-ratio of 2:1, pH-stability and addition of activators are important for the determination of galactokinase activity in the supernatant fraction of pig liver. 3. Using the optimized method galactokinase activity of pig liver in dependence on age, with particular reference to the perinatal period, was determined. 4. Galactokinase activity of liver of newborn piglets is 7 times that of adult pigs. In the suckling period the activity remains relatively constant at this high level and decreases remarkably immediately after weaning. 5. Galactokinase of liver of newborn piglets differs in kinetic properties (lower Km of ATP, higher maximal reaction velocity) from the enzyme of adult pigs, which is still insufficient to make sure the existence of two different forms of the enzyme.

The influence of carrageenan (0.1, 0.05 and 0.01%), galactose (1 X 10(-1), 1 X 10(-2) and 5 X 10(-2) mol/l) and 2,4-dinitrophenol (5 X 10(-4), 1 X 10(-4), 5 X 10(-5) and 1 X 10(-5) mol/l) on the phagocytosis of Staphylococcus aureus SG 511 and E. coli 04/K31 by polymorphonuclear neutrophil granulocytes was studied. At the same time the capacity of the germs to grow and multiply in presence of different concentrations of the analysed compounds was tested. The phagocytosis of Staphylococcus aureus SG 511 was not inhibited by carrageenan, galactose and 2,4-dinitrophenol, but an influence on the phagocytosis of E. coli 04/K31 could be clearly demonstrated (0.1 and 0.05% carrageenan, 1 X 10(-1) mol/l galactose, 5 X 10(-4) and 1 X 10(-4) mol/l 2,4-dinitrophenol). The multiplication of Staphylococci and E. coli was not influenced by all three compounds.