Multicomponent analysis of fermentation growth media using the electronic tongue (ET).

A potentiometric electronic tongue (ET) consisting of eight cross-sensitive chemical sensors and a standard pH electrode has been applied for analysis of simulated fermentation solutions typical for fermentation processes with Aspergillus niger. The electronic tongue has been found capable of simultaneous determination of ammonium, citrate and oxalate in complex media with good precision (typical error within 8%). The system preserved high sensitivity to the targeted substances also in the presence of sodium azide, which is commonly used for suppressing microbial activity in real-world fermentation samples. Sensor performance was fast and reproducible which promises well for routine application of the electronic tongue for fermentation process monitoring.

Chemometric evaluation of urinary steroid profiles in doping control.

Ten endogenous steroid hormones and metabolites were determined according to the screening procedure for anabolic steroids in spot urine samples from 105 healthy young male athletes (control samples) and 23 males that tested positive for anabolic steroids in the doping control (positive samples). The GC-MS peak areas for each sample were normalized to total area. Multivariate data analysis by Partial Least Square Regression (PLSR) and using a coded Y-variable (positive samples: +1 and control samples -1) allows projection of the most systematic profile structures into a 2D plot revealing a clear distinction between the control and misuser groups. The most important determinants of the location in the loading plot were the ratios of testosterone to epitestosterone and androsterone to etiocholanolone. The ratio between 11-beta-hydroxyandrosterone and 11-beta-hydroxy-etiocholanolone was less important, in accordance with the fact that anabolic-androgenic steroid intake primarily affects the excretion of testosterone from the testis and to a much lesser degree adrenal steroid genesis. We present a preliminary validation of this model (PLS1-DISCRIM) for analysing steroid profiles in doping control samples from several categories of athletes, some of which are suspected for drug misuse, and results from a one dose excretion study in healthy volunteers. Our findings suggest that use of multivariate PLS-regression may give valuable information about anabolic androgenic steroid misuse in sport. When appropriately calibrated, this methodology may delineate drug misusers directly from the screening procedure for anabolic steroids in spot urine tests.

Endocytosis and degradation of bovine apo- and holo-lactoferrin by isolated rat hepatocytes are mediated by recycling calcium-dependent binding sites.

We characterized endocytosis of iron-saturated (holo) and iron-depleted (apo) 125I-labeled bovine lactoferrin (Lf) by isolated rat hepatocytes. Hepatocytes ingested both Lf forms--determined by EGTA/dextran sulfate removal of surface-bound Lf--at maximal endocytic rates of 1.85 and 1.52 fmol cell-1 min-1 for 125I-apo-Lf and 125I-holo-Lf, respectively. First-order endocytic rate constants (37 degrees C) for 125I-apo-Lf and 125I-holo-Lf were 0.276 and 0.292 min-1, respectively. Regardless of Lf's iron content, hyperosmotic media (approximately 500 mmol/kg) inhibited Lf uptake by approximately 90%, indicating endocytosis of both Lf forms was primarily clathrin-dependent. Endocytosis of both Lf forms was not altered significantly in the presence of excess iron chelator desferrioxamine or rat holo-transferrin, or by cycloheximide treatment. Fluorescein isothiocyanate- and cyclohexanedione-modified Lf competed fully with native Lf for binding and endocytosis, indicating that, unlike human Lf, modification of lysine or arginine residues does not block the interaction of bovine Lf with cells. After binding Lf at 4 degrees C, cells at 37 degrees C internalized approximately 90% of Lf bound to Ca(2+)-dependent sites but not Lf bound to Ca(2+)-independent sites. Following uptake, hepatocytes released acid-soluble (degraded) products of 125I-Lf biphasically at 37 degrees C, an initial rapid phase within the first 20 min--more pronounced with 125I-holo-Lf--followed by a sustained linear release of 298 and 355 molecule equiv cell-1 min-1 for 125I-apo-Lf and 125I-holo-Lf, respectively. At 4 degrees C, both digitonin-permeabilized and intact cells bound approximately 1.1 x 10(6) 125I-Lf molecules to Ca(2+)-dependent sites per cell, indicating that hepatocytes do not contain a sizeable intracellular pool of these sites. Moreover, cells retained > 70% of Ca(2+)-dependent sites on the surface during sustained Lf endocytosis. Thus, these Lf binding sites recycle during endocytosis at an estimated 4-5 min/circuit.

Granulocyte elastase, beta-thromboglobulin, and C3d during acetate or bicarbonate hemodialysis with Hemophan compared to a cellulose acetate membrane.

Twenty-two patients were dialysed in a cross-over design using Hemophan or cellulose acetate membranes. The dialysate buffer was acetate (n = 12) or bicarbonate (n = 10). Blood was sampled at 0, 15, 60 and 180 min and mean values were adjusted for changes in total protein in each sample. At 15 min during dialysis a decrease in leukocytes and platelets occurred with both membranes, irrespective of the buffer (Wilcoxon, p less than 0.006). During dialysis, increases were found in granulocyte elastase inhibitor complex (E- alpha 1-PI), beta-thromboglobulin and C3d. beta 2-microglobulin was not significantly changed in blood after dialysis with Hemophan or cellulose acetate membranes with bicarbonate buffer. Side effects were more pronounced at 180 min during dialysis with bicarbonate in patients using cellulose acetate than with Hemophan (p = 0.021, n = 8). Hemophan seemed to be more favourable than cellulose acetate membranes in regard to leukopenia and E- alpha 1-PI. The dialysate buffer may also alter membrane biocompatibility.

Binding and endocytosis of apo- and holo-lactoferrin by isolated rat hepatocytes.

We characterized binding and endocytosis of 125I-bovine lactoferrin by isolated rat hepatocytes. Iron-depleted (apo-Lf), approximately 30% saturated (Lf), and iron-saturated (holo-Lf) lactoferrin were used. At 4 degrees C, cells bound 125I-apo-Lf and 125I-holo-Lf with nearly identical apparent first order kinetics (t1/2 = approximately 42 min). Holo-Lf and apo-Lf competed with each other for binding. Hepatocytes bound lactoferrin optimally at pH greater than or equal to 7 but poorly at pH less than or equal to 6. Ca2+ (greater than or equal to 100 microM) enhanced Lf binding to cells, and holo-Lf remained monomeric with Ca2+ present as determined by gel filtration chromatography. With Ca2+, cells exhibited approximately 10(6) high affinity sites (Kd approximately 20 nM) and approximately 10(7) low affinity sites (Kd approximately 700 nM) for both apo- and holo-Lf. Without Ca2+, cells bound 125I-holo-Lf by the low affinity component only. EGTA and dextran sulfate together released greater than or equal to 90% 125I-Lf prebound at 4 degrees C, but individually removed separate populations of surface-bound 125I-Lf. Cells bound 125I-Lf in a Ca(2+)-dependent manner with dextran sulfate present. We conclude that the high affinity but not the low affinity sites require Ca2+; only the low affinity sites are dextran sulfate-sensitive. Neither transferrin nor asialo-orosomucoid blocked lactoferrin binding to hepatocytes. Some cationic proteins but not others inhibited lactoferrin binding. At 37 degrees C, hepatocytes endocytosed 125I-apo-Lf and 125I-holo-Lf similarly, and hyperosmolality (greater than 500 mmol/kg) blocked uptake by approximately 90%. These data support the proposal that hepatocytes regulate blood lactoferrin concentration by receptor-mediated endocytosis.

A multivariate pattern recognition study of risk-factors indicating postoperative thromboembolism despite low-dose heparin in major abdominal surgery.

The object of the present investigation was to identify those who, among high-risk patients, would "break through" low-dose heparin prophylaxis and develop thromboembolism after major abdominal surgery. Twenty-nine variables (clinical characteristics, pre- and postoperative coagulation and fibrinolytic factors) from 19 patients with and 26 patients without thromboembolism were analyzed by means of a multivariate supervised pattern recognition technique (SIMCA). We found no statistically significant difference between patients with and without thromboembolism. Thus, in the studied group of high-risk patients it was not possible to identify a predictive index for selection of individual patients liable to develop postoperative thromboembolism despite low-dose heparin prophylaxis in major abdominal surgery.

Ulcer recurrence after parietal cell vagotomy for duodenal ulcer. A multivariate pattern recognition study.

The object of the study was to identify individual high-risk patients with regard to ulcer recurrence after parietal cell vagotomy for duodenal ulcer. The study comprises a multivariate analysis of 14 variables (age, sex, duration of symptoms, site of ulcer, pre- and post-operative acid secretion) for 37 patients with and 111 patients without recurrence 5-11 years after parietal cell vagotomy for duodenal ulcer. The data were analyzed using a supervised pattern recognition technique, SIMCA (Soft Independent Modeling of Class Analogy), which analyzes complex data as geometrical elements in a multidimensional space. We found no statistical difference between the patients with and without ulcer recurrence. Thus, no predictive value for the selection of patients liable to develop recurrent ulcer after parietal cell vagotomy was contained in the variables generally registered in current surgery.