Age-dependent changes of proinflammatory cytokine production by porcine peripheral blood phagocytes

The aim of the present study was to determine postnatal ontogeny of proinflammatory cytokines IL-1beta, IL-8 and TNF-alpha production by in vitro stimulated porcine blood leukocytes. Four age categories of pigs were chosen. Cytokine production was determined using intracellular flow cytometry. It was found that IL-8 and TNF-alpha production by blood monocytes significantly increased during the postnatal period while production of IL-1beta remained unchanged. In blood neutrophils, the IL-8 production increased only during the postnatal period, while the levels of TNF-alpha and IL-1beta were undetectable during the whole postnatal period. Generally, the most intensive changes in cytokine production occurred before weaning. The production of low levels of cytokines by monocytes and neutrophils from young pigs was not caused by a delayed cytokine response because the cytokine production after 8-h stimulation was lower than that after 4-h stimulation in all age categories. The ontogenetical changes showed the same trends when two different stimulators (LPS, heat-inactivated E. coli) were used, suggesting that the ontogenetical changes are not caused by a simple defect in one signalling pathway, but it is probably a more complex process. No differences in cytokine production between the whole blood and the isolated cells supplemented with newborn or adult serum were found. Thus the ability of newborn monocytes and neutrophils to produce proinflammatory cytokines was not decreased due to the influence of composition of the microenvironment, where the cells were present. In conclusion, the ability of porcine blood leukocytes to produce cytokines develops during postnatal life.     

Effect of varied soil nitrogen supply on growth and nutrient uptake of young Norway spruce plants grown in a shaded environment

To understand the effect of increased soil N supply on tree growth and nutrient uptake, three-year-old Norway spruce seedlings were grown in pots on low-nutrient mineral forest soil supplemented with N in mineral or organic form. Outdoor shaded growth conditions were used, to test the hypothesis that shaded plants are particularly susceptible to high soil N supply. Plants were harvested eleven months after planting. Shoot growth was not affected by the N supply, but N concentrations in needles and roots were increased in plants supplied with mineral N (150 or 300 mg N [kg soil]^—1). Root growth was drastically reduced and root/shoot ratios were decreased in plants with higher N uptake. A high supply of mineral N to soil also decreased the concentrations of other essential elements (P, K) in the needles and thus had effects on plant growth which may impair the stress resistance of trees. Organic N in the form of keratin (150 mg N [kg soil]^—1) did not influence plant growth significantly. The adverse effects of high mineral N supply were particularly pronounced under shaded conditions in comparison to results from other experiments using higher light intensity and temperature conditions.     

Aluminium and lead tolerance in ectomycorrhizal fungi

Al and Pb tolerance of different ectomycorrhizal fungi was tested in Petri dishes divided into three compartments. One compartment contained nutrient solution supplemented with Al or Pb but without P to avoid precipitation of Al or Pb phosphates. Phosphate was supplied in the agar compartment while a third compartment served as control containing nutrient solution without P. The tested ectomycorrhizal fungal species and isolates varied greatly in their Al and Pb tolerance. A more than 50% growth reduction was observed at 2000 μM Al in Paxillus involutus MAI but already at 200 μM in P. involutus NAU and P. involutus 533. In contrast, growth of Pisolithus tinctorius 956 appeared to be stimulated by Al. Al tolerance therefore decreased in the following order: P. tinctorius 956 > P. involutus MAI > P. involutus NAU, P. involutus 533. To test the effect of agar on Al tolerance, P. involutus MAI was grown with agar in all three compartments. In presence of agar, even 2000 μM Al had no effect on growth. Lead at concentrations of 1 and 10 μM Pb had no significant effect on fungal growth. A more than 50% decrease of mycelial surface area occurred at 500 μM Pb in P. involutus MAI and P. involutus 533 and at 100 μM Pb in P. involutus NAU and P. tinctorius 956. Pb tolerance therefore decreased in the following order: P. involutus MAI, P. involutus 533 > P. involutus NAU, P. tinctorius 956. Divided Petri plates appear to be a valuable tool to test metal tolerance of fungi as complexation of the metals by agar or precipitation with P are avoided. The results indicate that some ectomycorrhizal fungi may be more sensitive to Al and Pb than previously reported.     

Growth characteristics of porcine chlamydial strains in different cell culture systems and comparison with ovine and avian chlamydial strains

Porcine Chlamydiaceae were cultivated under various culture conditions and we compared their growth characteristics with those of ruminant and avian strains. The combination of centrifugation assisted cell culture infection and cycloheximide treatment of Vero cell coverslip cultures provided the highest inclusion numbers with all chlamydial strains. Interestingly, the use of Iscove's modified Dulbecco's medium instead of Eagle's minimal essential medium significantly increased Chlamydia suis inclusion counts. C. suis and Chlamydophila pecorum inclusion numbers were markedly increased in CaCo cells, compared with Vero cells. This accelerated growth of porcine Chlamydiaceae under certain cultivation conditions may be helpful for the propagation of low chlamydial numbers or for their isolation from field samples. The intracellular distribution of porcine Chlamydiaceae in polarised CaCo cells clearly demonstrated differences between the chlamydial strains: C. pecorum 1710S inclusions were predominantly localised in the apical cytoplasm, C. suis S45 inclusions, however, were mostly situated in lower cytoplasmatic compartments. These findings might reflect biological differences in vivo.     

Inhibited and uninhibited infants "grown up": adult amygdalar response to novelty

Infants with an inhibited temperament tend to develop into children who avoid people, objects, and situations that are novel or unfamiliar, whereas uninhibited children spontaneously approach novel persons, objects, and situations. Behavioral and physiological features of these two temperamental categories are moderately stable from infancy into early adolescence and have been hypothesized to be due, in part, to variation in amygdalar responses to novelty. We found that adults who had been categorized in the second year of life as inhibited, compared with those previously categorized as uninhibited, showed greater functional MRI signal response within the amygdala to novel versus familiar faces.     

Mutations in the pericentrin (PCNT) gene cause primordial dwarfism

Fundamental processes influencing human growth can be revealed by studying extreme short stature. Using genetic linkage analysis, we find that biallelic loss-of-function mutations in the centrosomal pericentrin (PCNT) gene on chromosome 21q22.3 cause microcephalic osteodysplastic primordial dwarfism type II (MOPD II) in 25 patients. Adults with this rare inherited condition have an average height of 100 centimeters and a brain size comparable to that of a 3-month-old baby, but are of near-normal intelligence. Absence of PCNT results in disorganized mitotic spindles and missegregation of chromosomes. Mutations in related genes are known to cause primary microcephaly (MCPH1, CDK5RAP2, ASPM, and CENPJ).     

Efficient inhibition of the Alzheimer's disease beta-secretase by membrane targeting

beta-Secretase plays a critical role in beta-amyloid formation and thus provides a therapeutic target for Alzheimer's disease. Inhibitor design has usually focused on active-site binding, neglecting the subcellular localization of active enzyme. We have addressed this issue by synthesizing a membrane-anchored version of a beta-secretase transition-state inhibitor by linking it to a sterol moiety. Thus, we targeted the inhibitor to active beta-secretase found in endosomes and also reduced the dimensionality of the inhibitor, increasing its local membrane concentration. This inhibitor reduced enzyme activity much more efficiently than did the free inhibitor in cultured cells and in vivo. In addition to effectively targeting beta-secretase, this strategy could also be used in designing potent drugs against other membrane protein targets.