Persistent changes in arterial blood gases in fetal sheep

Two anaesthetic protocols were compared using pregnant sheep. In both groups of animals, anaesthesia was induced using an intravenous (i.v.) injection of diazepam and ketamine. The ewes were then intubated for positive pressure ventilation using 0.8 L/min of nitrous oxide and 2 L/min oxygen with 1.1-1.8% halothane. If the ewe showed any signs of awakening, one of two protocols was followed. First, the halothane concentration was increased to 2-3% until the ewe was completely anaesthetized. Second, the halothane concentration was not altered, but the ewe was given doses of i.v. diazepam (0.1 mg/kg) and ketamine (1 mg/kg) until again completely anaesthetized. At the completion of surgery, maternal recovery was rapid and similar between the two groups. However, five days after surgery, the fetal arterial Po(2) and oxygen content of the fetuses receiving additional halothane (1.9 +/- 0.2 kPa and 4.4 +/- 1.0 mL/100 mL) were statistically significantly depressed when compared with the fetuses receiving additional diazepam and ketamine (2.9 +/- 0.1 kPa and 7.0 +/- 0.5 mL/100 mL). These results led us to conclude that certain anaesthetic protocols, in spite of good maternal recovery, can lead to deleterious effects upon the fetus that persist for at least five days after surgery.     

Predictors of dementia, the construction of classification trees

In order to identify persons who are at risk for dementia in an early phase, two classification trees were developed. Data were used from the Longitudinal Aging Study Amsterdam (LASA). The prevalence of dementia in the whole sample was 4.0%. In the first tree age seemed to be the strongest predictor, with an increased risk for persons older than 75. In this group the positive predictive value reached a maximum of 33.3% when the persons had memory complaints and a score on the Mini Mental State Examination (MMSE) <24. In a second classification tree, age was excluded as a predictor because of high association with the other potential predictors. In this tree functional limitations seemed the strongest predictor. In the group of persons with at least one functional limitations, the positive predictive value reached a maximum of 28.8% when the persons had memory complaints and a score <24 on the MMSE. In persons without memory complaints, persons with cardiovascular diseases or diabetes were at increased risk of dementia. Further research is necessary before these classification trees can be implemented in general health care.     

Quantification of viability in organotypic multicellular spheroids of human malignant glioma using lactate dehydrogenase activity: a rapid and reliable automated assay.

Organotypic spheroids from malignant glioma resemble the biological complexity of the original tumor and are therefore appealing to study anticancer drug responses. Accurate and reproducible quantification of response effect has been lacking to determine drug responses in this three-dimensional tumor model. Lactate dehydrogenase (LDH) activity was demonstrated in cryostat sections of spheroids using the tetrazolium salt method. Calibrated digital image acquisition of the stained cryostat sections enables quantification of LDH activity. Fully automated image cytometry reliably demarcates LDH-active and LDH-inactive tissue areas by thresholding at specific absorbance values. The viability index (VI) was calculated as ratio of LDH-active areas and total spheroid tissue areas. Duplicate staining and processing on the same tissue showed good correlation and therefore reproducibility. Sodium azide incubation of spheroids induced reduction in VI to almost zero. We conclude that quantification of viability in cryostat sections of organotypic multicellular spheroids from malignant glioma can be performed reliably and reproducibly with this approach.     

Selective grazing by adults and larvae of the zebra mussel (Dreissena polymorpha): application of flow cytometry to natural seston

1. Selective grazing of adults and larvae of the zebra mussel (Dreissena polymorpha) on phytoplankton and detritus from both laboratory cultures and natural seston was quantified using flow cytometry. 2. Mean clearance rate of adult zebra mussels was higher on a mixture of the green alga Scenedesmus and the cyanobacterium Microcystis than when Scenedesmus was offered as single food, suggesting selective feeding by the mussels. 3. Feeding on lake seston both adults and larvae showed a higher clearance rate on phytoplankton than on detritus particles, suggesting that zebra mussels select for phytoplankton. Furthermore, it was noted that adults preferred seston particles in the 0–1 and 30–100 μm size ranges. 4. In our study, zebra mussels did not discriminate against cyanobacteria, and our results indicate that they may even ingest them preferentially.     

Serum amyloid-A (SAA) and haptoglobin (Hp) plasma concentrations in newborn calves

The concentrations of 2 major bovine acute-phase proteins, haptoglobin (Hp) and serum amyloid-A (SAA), were measured in plasma obtained shortly after birth from 22 healthy calves. In a separate group of diseased calves (n = 8), Hp and SAA concentrations were measured to determine whether newborn calves (up to 4 d old) are able to produce SAA and Hp. In blood samples taken directly after birth, the Hp plasma concentrations were all below the limit of detection. The mean SAA concentration was independent of weight (r = 0.063), degree of acidosis (r = -0.125), sex (p > 0.05), and were not different in calves born after different types of obstetrical help (p > 0.05). In the group of diseased calves, an increased Hp concentration was measured in only 2 of 8 animals, whereas the mean SAA concentration was significantly higher (p < 0.05) than in the healthy newborn calves. These data suggest that prenatal stress due to parturition does not form a stimulus for the production of acute-phase proteins in the fetal calf. The low Hp plasma concentrations might indicate that either it takes a few days to establish a detectable concentration of this protein, or that Hp production is not fully developed in newborn calves.     

COMPETITION BETWEEN A PROCHLOROPHYTE AND A CYANOBACTERIUM UNDER VARIOUS PHOSPHORUS REGIMES: COMPARISON WITH THE DROOP MODEL

The ecophysiology and competitive behavior of the prochlorophyte Prochlorothrix hollandica Burger-Wiersma, Stal et Mur, and the cyanobacterium Planktothrix agardhii Anagn. et Kom. were investigated in phosphorus-limited continuous cultures. When the species were exposed to successive saturating pulses of P, the maximal P uptake rate decreased linearly with an increase in the P cell quota. Prochlorothrix had a higher maximal P uptake rate, a lower half-saturation constant for P uptake, higher maximal cell quota for P, and slightly lower minimal cell quota for P than Planktothrix. These data indicate that Prochlorothrix is an affinity and storage strategist, at least when compared to Planktothrix. On the other hand, Prochlorothrix had a lower maximal growth rate than Planktothrix. On the basis of these ecophysiological parameters, we developed a Droop model to predict the time course and outcome of competition under various P regimes. The model predictions were in line with the results of competition experiments under three different P-limited conditions (continuous P supply, 4-day pulse period, 12-day pulse period). Prochlorothrix competitively displaced Planktothrix under both a constant and a pulsed P supply, and competitive displacement of Planktothrix was slowest in the experiment with a 12-day pulse period. In the pulsed experiments, the mean filament lengths and chl a fluorescence of the species oscillated at the same frequency as the pulse additions. In contrast to the Droop model, the predictions of the Monod model were not in line with the outcome of the competition experiments. Our results demonstrate that Prochlorothrix is a very good competitor for P and that the time course and outcome of competition for P in a variable environment can be predicted on the basis of the P uptake and storage characteristics of the species.     

Cardiotocography in cows: A method for monitoring calves during delivery

Fetal ECG-electrodes and an intrauterine catheter were inserted into 15 Holstein-Friesian cows during their first stage of labor to make a cardiotocogram. Simultaneously, fetal heart rate and intrauterine pressure were recorded until completion of fetal expulsion. Immediatly post partum the viability of the calf was assessed by clinical evaluation and measurements of blood pH, base excess (BE) and pCO(2). Fetal heart rate patterns and their changes were evaluated according to standards used in human medicine. Basal fetal heart rate (90 to 120) in 10 calves gradually increased towards the end of parturition, with a marked loss of variability. Decelerations, coinciding with periods of increased intrauterine pressure, occurred in all cows. When decelerations occurred beyond the end of a contraction the calf was born in a poor condition. Accelerations were only recorded in two cows. Many of the fetal heart rate changes observed were similar to those which in human cardiotocography are considered to be signs of fetal distress. Further investigation is needed to establish the predictive value of fetal heart rate patterns in cows, which would be a welcome addition to improved fetal diagnostics during parturition.     

The chimpanzee major histocompatibility complex class II DR subregion contains an unexpectedly high number of beta-chain genes

The major histocompatibility complex (MHC) class II DR subregion of the chimpanzee was studied by restriction fragment length polymorphism (RFLP) analysis. Genomic DNA obtained from a panel of 94 chimpanzees was digested with the restriction enzyme Taq I and hybridized with an HLA-DR probe specific for the 3' untranslated (UT) region. Such a screening revealed the existence of 14 distinct DRB/Taq I gene-associated fragments allowing the definition of 11 haplotypes. Segregation studies proved that the number of chimpanzee class II DRB/Taq I fragments is not constant and varies from three to six depending on the haplotype. Comparison of these data with a human reference panel manifested that some MHC DRB/Taq I fragments are shared by man and chimpanzee. Moreover, the number of HLA-DRB/Taq I gene-associated fragments detected in a panel of homozygous typing cells varies from one to three and corresponds with the number of HLA-DRB genes present for most haplotypes. However, a discrepancy is observed for the HLA-DR4,-DR7, and -DR9 haplotypes since a fourth HLA-DRB pseudogene present within these haplotypes lacks its 3' UT region and thus is not detected with the probe used. These results suggest that chimpanzees have a higher maximum number of DRB genes per haplotype than man. As a consequence, some chimpanzee haplotypes must show a dissimilar organization of the MHC DR subregion compared to their human equivalents. The implications of these findings are discussed in the context of the trans-species theory of MHC polymorphism. Address correspondence and offprint requests to: R. E. Bontrop.