Automated fluorimetry in quality control of pasteurized and ultra-high temperature-treated starch soup

Automated monitoring of the microbiological quality of heat-processed foods by the resazurin reduction test was applied to microtitration plate incubator-fluorimeter technology. The appearance and disappearance of the fluorescing peak of resorufin was monitored on microtitration trays. Pasteurized or ultra-high temperature-treated starch-based soup was used as the model food system. Bacillus subtilis spores (ultra-high temperature treatment) and vegetative cells of Enterococcus faecalis (pasteurization) were inoculated into the soup before the heat treatment at levels which resulted in some survival. The timing of appearance of maximum fluorescence correlated with the number of bacteria in pre-incubated samples. Automated resazurin-reduction fluorimetry was compared with conventional plating, turbidometry and microcolony count by the direct epifluorescent filter technique. The results of the resazurin test correlated well with those of all the other methods tested. Fluorimetry had the advantage that the results could be read within 1–5h and the reproducibility was superior to the other methods.     

Fluorescence flow cytometry of human leukocytes in the detection of LDL receptor defects in the differential diagnosis of hypercholesterolemia

A flow-cytometric method with fluorescence-labeled monoclonal antibodies (MABs) against the low density lipoprotein (LDL) receptor (C7A MAB) or 3,3'-dioctadecylindocarbocyanin-iodide (DiI) LDL has been developed that allows the quantification of LDL receptors on leukocytes and the identification of patients with familial hypercholesterolemia (FH) within 48 hours. Leukocytes were isolated from 10 mL anticoagulated blood by density gradient centrifugation. To induce maximal expression of LDL receptors, mononuclear cells were preincubated with either phytohemagglutinine (PHA) or lipoprotein-deficient serum (LPDS). LPDS-treated monocytes provided a more homogeneous cell population with regard to LDL receptor activity than did the PHA-treated lymphocytes; they also provided a greater discrimination between the fluorescence of the receptor probes and cellular autofluorescence. The C7A MAB was able to compete for DiI LDL binding by about 40%. In competition with unlabeled LDL, DiI LDL revealed linear binding, indicating an affinity similar to native LDL. The binding characteristics of DiI LDL were also similar to 125I-LDL binding. LDL isolated from familial defective apolipoprotein B-100 was not able to compete for DiI LDL binding on monocytes, whereas native LDL reduced it by about 80%. In monocytes from FH heterozygous patients, the cellular mean fluorescence using either C7A MAB or DiI LDL at 4 degrees C was 30% to 70%; in FH homozygotes, cellular mean fluorescence was less than 20% of that in monocytes from normal individuals. In patients with familial defective apolipoprotein B-100 antibody binding was normal, but one patient's own LDL failed to compete with normal DiI LDL for 4 degrees C binding on U937 test monocytes. Patient monocytes having internalization defects showed normal 4 degrees C DiI LDL binding, but at 20 degrees C cell-associated fluorescence was reduced by about 40%. In our study 384 hypercholesterolemic patients (preselected according to serum cholesterol levels, clinical symptoms, and family history) were analyzed for LDL receptor expression using the C7A MAB-based assay. In 71.8% of the patients with cholesterol levels higher than 300 mg/dL, an LDL receptor deficiency was observed. Apolipoprotein E isoforms and lipoprotein[a] were found to be independent from the LDL receptor status. In some patients with high cholesterol levels but normal LDL receptor expression with the C7A MAB assay, LDL receptor defects could be diagnosed when either reduced binding or internalization of DiI LDL or familial defective apolipoprotein B-100 was detected.(ABSTRACT TRUNCATED AT 400 WORDS)     

The femur-tibia control system of stick insects--a model system for the study of the neural basis of joint control

In a form of top-down analysis, the femur-tibia control system of stick insects is investigated. Open-loop experiments show that it is mainly velocity-sensitive with an extremely low velocity-threshold, that it possesses a very high gain and that it has only a very small phase reserve and thus works close to instability. The closed-loop system generates catalepsy. The system consists of a single sense organ with approximately 80 sense cells with known characteristics, a small number of interneurones (mainly non-spiking ones) and a small number of motor neurones. The characteristics of the whole system can quantitatively be attributed to the characteristics of its elements. The gain of the loop is state-dependent and the system is 'switched off' during active movements and replaced by a control system with different attributes. It is discussed that most of the characteristics of this system are, at least qualitatively, similar to joint control systems in other animals. Because the described system can be more easily investigated than other systems (especially in vertebrates) it can serve as a model against which more complicated joint control loops may be compared.     

Nontuberculous mycobacteriosis as a complication of the Swyer-James syndrome

The chest radiograph of a 35-year-old man with fatigue, exertional dyspnoea and haemoptyses revealed a cavity in the left upper lobe and a shrunken left lung with radiolucency greater than that on the right. Acid-fast rods in sputum were identified as Mycobacterium kansasii on culture. Scintigraphy showed a 9% residual perfusion on the left and abnormal ventilation, compatible with Swyer-James syndrome. This had favoured the development of a mycobacterial infection. There was also a decrease in ciliary function (rate of 4-7 Hz, normal: 10-11). Treatment, begun when tuberculosis had been suspected, was after sensitivity tests changed to a combination of rifampicin (600 mg), ethambutol (1600 mg) and protionamide (500 mg) daily. There was marked regression of the findings within 4 weeks, but treatment was prematurely stopped after 11 months. Two years later there was a recurrence which again responded well to the same drug regimen with additional sulphamethoxazole (1600 mg/d).     

Characterisation of sub-100 nm-MOS-transistors processed by optical lithography and a sidewall-etchback technique

This paper describes the fabrication of NMOS-transistors with a geometric gate length of down to 50 nm using conventional optical lithography and a modified sidewall-etchback process. Based on measurements the transistors are characterised and their device parameters are compared to simulations. Finally the procedures for further optimisation of the process will be explained.     

Nature of terpenoids of aTaxodioxylon gypsaceum from the fossil forest of Dunarobba (Italy)

A specimen fromt he fossil forest of Dunarobba (Central Italy) attributed toTaxodioxylon gypsaceum was chemically analyzed particularly for its terpene content. Main chemical constituents show variations related to ageing and carbonization process, while terpene content exhibits a drop of mono- and sesquiterpenes in favour of diterpenes and higher MW compounds. All components are exclusively hydrocarbons and represent alicyclic or partially aromatic structures. The identified families include abietane, phyllocladane, norabietane and norpimarane among diterpenes and traces of cadinane and selinane among sesquiterpenes.