The production of hematopoietic growth factors by endothelial accessory cells

Monocytes are known to produce both hematopoietic growth factors and other factors, monokines, which do not directly stimulate hematopoiesis. Monokines such as interleukin-1 (IL-1) and tumor necrosis factor (TNF) may indirectly stimulate mesenchymal cells to produce hematopoietic growth factors. The identity of all the factors produced by monocytes or mesenchymal cells has not been established because of overlapping activities on biologic assay. The purpose of this study was to identify the individual growth factors produced by endothelial cells before and after stimulation with various monokines. We prepared conditioned media and extracted RNA from endothelial cells before and after stimulation with monokines. The results show that immortalized endothelial cells produce maximal detectable amounts of granulocyte-macrophage colony-stimulating factor (GM-CSF) constitutively. In contrast, GM-CSF production by primary endothelial cells requires induction with either IL-1 or TNF.     

Interactions between Males of the Lizard Liolaemus tenuis: Roles of Familiarity and Memory

For territorial organisms, recognition of familiar individuals can reduce the frequency and intensity of aggressive encounters (‘dear enemy’ phenomenon), stabilize social systems, and reduce the cost of territory maintenance. Here, we investigated the behavioural events displayed during contests between familiar and unfamiliar individuals in the lizard Liolaemus tenuis (Liolaemidae), a species in which males are territorial. The behaviours recorded were attack, warning, evasion, and submission, and the latencies to the first aggressive (attack or warning) behaviour. Additionally, we assessed the ability of individuals to remember a familiar conspecific after a period without social interaction. Individual males reduced and delayed aggressive behaviour directed towards socially familiar individuals compared with unfamiliar ones. These results suggest that males distinguished between familiar and unfamiliar conspecific males and are in agreement with the ‘dear enemy’ phenomenon. Other behaviours were similar in the contests between familiar and unfamiliar individuals. Recognition of familiar conspecifics was lost after 20 d without social interactions. This may be relevant for interactions with floater males or with neighbours that lose their territory and subsequently attempt to fight for their ex‐neighbour's territory.     

Inhibition of hexokinase activity by a fructose 2,6-bisphosphate-dependent cytosolic protein from liver

Mammalian and yeast hexokinases were found to be reversibly inhibited by fructose 2,6-bisphosphate, an effect requiring the presence of a cytosolic protein factor. Experimental evidence suggests that this factor (inhibitor) is a regulatory protein, the interactions of which with hexokinases are modulated by fructose 2,6-bisphosphate. The Vmax of hexokinase D was decreased, and no changes on other kinetic parameters were observed. The inhibitor was present in fresh liver cytosol filtered through Sephadex G-25 and was partially isolated by negative absorption on DEAE-cellulose followed by ammonium sulfate fractionation. The inhibitor was also present in brain and kidney, but not in muscle. A molecular mass of 200,000 was determined by gel filtration. The inhibition was dependent on the concentrations of both the inhibitory protein and fructose 2,6-bisphosphate. No delay in fructose 2,6-bisphosphate inhibition was observed. Several other hexose phosphates were tested and were not effective. In the presence of amounts of inhibitor sufficient to produce complete inhibition of hexokinase D, the concentration of fructose 2,6-bisphosphate required to produce 50% inhibition was about 0.5 microM. The inhibitor was unstable and was stabilized by the presence of fructose 2,6-bisphosphate.     

Phosphofructokinase is responsible for the fructose 2,6-bisphosphate inhibition of hexokinase in tissue extracts

Mammalian and yeast hexokinases were reported to be reversibly inhibited by fructose 2,6-bisphosphate in the presence of cytosolic proteins (H. Niemeyer, C. Cerpa, and E. Rabajille (1987) Arch. Biochem. Biophys. 257, 17-26). Reinvestigation of this finding using a radioassay with [14C]glucose as substrate showed no effect of fructose 2,6-bisphosphate on hexokinase activity of rat liver cytosols. Detailed reexamination of the spectrophotometric assay resulted in the observation that the fructose 2,6-bisphosphate-dependent inhibition was a function of the cytosolic phosphoglucose isomerase and phosphofructokinase activities compared to the amount of glucose-6-phosphate dehydrogenase used as auxiliary enzyme. The diminution or loss of the fructose 2,6-bisphosphate-dependent inhibition produced in aged cytosols was restored by addition of crystalline muscle phosphofructokinase, as well as by decreasing the amount of glucose-6-phosphate dehydrogenase in the assay. When phosphoglucose isomerase, phosphofructokinase, and hexokinase activities were separated by DEAE-chromatography of liver cytosol, no fructose 2,6-bisphosphate-dependent inhibition of hexokinase was found in any single fraction of the chromatogram. However, combination of fractions containing both phosphoglucose isomerase and phosphofructokinase displayed the fructose 2,6-bisphosphate-dependent inhibition on either endogenous hexokinase or added yeast hexokinase. From these results we conclude that the activation of phosphofructokinase elicited by fructose 2,6-bisphosphate is responsible for the hexokinase inhibition observed in the coupled spectrophotometric assay.     

Functional properties of palmaris longus muscles of rhesus monkeys transplanted as index finger flexors

This experiment with skeletal muscle autografts in monkeys was designed to retest previous findings that transplanted skeletal muscle can regenerate to a functional degree in primates without predenervation and to test a new hypothesis that increased functional demands on regenerated muscle grafts in monkeys may result in improved functional capacity of the grafts. Rhesus monkey index flexors were replaced with free palmaris longus muscle autografts with microneural anastomoses between the graft motor nerve and the severed profundus motor nerve. One monkey was taught selective index flexion before grafting and continued with this program after grafting to test the effect of training on the graft. Mature grafts were evaluated for in vivo contractile properties and by histology and histochemistry and were compared with a group of normal Rhesus palmaris longus muscles. The results reconfirm the capacity of nonpredenervated monkey skeletal muscle grafts to regenerate and to achieve some contractile ability and suggest that training of free muscle grafts may enhance recovery of their functional and structural properties.     

Agglutinierte Foraminiferen aus Graptolithen-Schwarzschiefern des Llanvirnium (Ordovizium) von Plettenberg im Sauerland (Nordrhein-Westfalen,NW-Deutschland)

The Plettenberger Bänderschiefer (Ordovician, Lower Llanvirnian, Zone ofDidymograptus bifidus (Hcall) from the type locality at Plettenberg (Sauerland, NW-Germany) yielded a foraminiferal fauna consisting of primitive agglutinated species (Amphitremoida, Bathysiphon,?Raibosammina, ?Thekammina, Thurammina). Forming one of the stratigraphically oldest European foraminiferal assemblages, these taxa were washed out of an unconsolidated graptolite siltstone and represent a pelagic, oxygen-depleted environment.     

Dominant Unc-37 Mutations Suppress the Movement Defect of a Homeodomain Mutation in Unc-4, a Neural Specificity Gene in Caenorhabditis Elegans

The unc-4 gene of Caenorhabditis elegans encodes a homeodomain protein that defines synaptic input to ventral cord motor neurons. unc-4 mutants are unable to crawl backward because VA motor neurons are miswired with synaptic connections normally reserved for their sister cells, the VB motor neurons. These changes in connectivity are not accompanied by any visible effects upon neuronal morphology, which suggests that unc-4 regulates synaptic specificity but not axonal guidance or outgrowth. In an effort to identify other genes in the unc-4 pathway, we have devised a selection scheme for rare mutations that suppress the Unc-4 phenotype. We have isolated four, dominant, extragenic, allele-specific suppressors of unc-4(e2322ts), a temperature sensitive allele with a point mutation in the unc-4 homeodomain. Our data indicate that these suppressors are gain-of-function mutations in the previously identified unc-37 gene. We show that the loss-of-function mutation unc-37(e262) phenocopies the Unc-4 movement defect but does not prevent unc-4 expression or alter VA motor neuron morphology. These findings suggest that unc-37 functions with unc-4 to specify synaptic input to the VA motor neurons. We propose that unc-37 may be regulated by unc-4. Alternatively, unc-37 may encode a gene product that interacts with the unc-4 homeodomain.     

Variations in the sulfide regime and the distribution of macrofauna in an intertidal flat in the North Sea

Sulfide concentrations were measured in the inner and outer Königshafen (Sylt, Wadden Sea) from November 1990 to December 1991 to assess the fluctuations of sulfide levels in natural tidal habitats. Three different areas were compared: (1) muddy sediment (2) fine-medium sand, and (3) a mussel bed. Other abiotic factors such as Eh, pH, temperature, grain size and organic content were measured. After assessment of the macrofaunal distribution, an attempt was made to relate the distribution to the sulfide concentrations in the benthic habitat. Sulfide concentrations varied between sites throughout the year with considerable differences ranging from below 5 μM (limit of detection) to about 1 mM (Oct. 1991). The faunal composition (Table 2) at the different sites hardly varied; it was always dominated by annelids: The cirratulidTharyx marioni was the most abundant species in the upper layers of all sites, where it occurred at low sulfide concentrations (<50 μM).Heteromastus filiformis was commonly found in the deeper sediment layers of the muddy site where it was regularly exposed to sulfide levels around 75 μM and peak concentrations in autumn up to 1 mM.Capitella capitata, Tubificoides pseudogaster andTubificoides benedii were very common in the upper sediment layers where sulfide levels were about 150 μM in autumn. These species also occurred, however, in the deeper layers with higher sulfide concentrations. These results document not only the wide annual range of sulfide concentrations at different depths in a tidal flat, they also emphasize that under natural conditions tidal flat annelids are exposed to considerable concentrations of hydrogen sulfide.     

Stability of hexokinases A, B and C and N-acetylglucosamine kinase in liver cells isolated from rats submitted to diabetes and several dietary conditions.

The effect of dietary and hormonal variations on the specific activities of hexokinase isoenzymes, N-acetylglucosamine kinase and pyruvate kinase isoenzymes in parenchymal and non-parenchymal liver cells was studied. Hexokinase D was markedly decreased in hepatocytes from animals fasted or fed on the carbohydrate-free diet as well as from diabetic rats, attaining a constant low level of about 17% of normal values. Pyruvate kinase L was also diminished in hepatocytes under the same experimental conditions. In contrast, the three high-affinity hexokinase isoenzymes A, B and C remained without variation in total amount or in their relative proportions in hepatocytes and non-parenchymal liver cells isolated from animals under the various conditions studied. N-Acetylglucosamine kinase activities also did not change either in parenchymal or in non-parenchymal liver cells under all conditions. The results are discussed in relation to the significance of N-acetylglucosamine kinase and the various hexokinase isoenzymes for the phosphorylation of glucose after dietary and hormonal manipulations.