cAMP mediates IL-1-induced lymphocyte penetration through endothelial monolayers

Endothelial cell incubated with IL-1 have been shown adhere more lymphocytes than nontreated endothelial cells. Here we demonstrate that IL-1 can also increase lymphocyte penetration through endothelial monolayers in vitro. IL-1 induced a transient increase in the number of lymphocytes penetrated through the endothelial monolayer into a filter in a time- and dose-dependent manner. This effect could be mimicked by increasing the cytosolic cAMP levels in the endothelial cells either by forskolin or dibutyryl-cAMP. Concomitantly we were able to show that IL-1 increased the cytosolic cAMP levels in endothelial cells. An inhibitor of adenylate cyclase, ddAdo, decreased both the IL-1-induced cAMP elevation and lymphocyte penetration. A protein kinase A inhibitor HA 1004 could inhibit the IL-1-induced lymphocyte penetration, where as protein kinase C (N-(2-guamidino-ethyl)-5-isoquinolinesyl foamide hydrocloride) and calcium-calmodulin (N-(6-aminohexyl)-5-chloro-1-naphthalensulfanamide) inhibitors had no effect. Adding dibutyryl-cGMP or calcium ionophore to the endothelial cells could not mimic IL-1-induced penetration and finally IL-1 did not induce PKC translocation in endothelial cells. These data support the view that IL-1 acts via cAMP as a second messenger in regard to lymphocyte penetration through endothelial cells. The above data demonstrate that IL-1-induced lymphocyte penetration through endothelial cells and that this IL-1-induced signal is transduced via cAMP in endothelial cells.     

A modified Kelsey-Sykes method for testing disinfectants with 2,3,5-triphenyltetrazolium chloride reduction as an indicator of bacterial growth

Different dilutions of chlorhexidine gluconate were tested by the Kelsey-Sykes procedure. The method was further modified on microtitration plates using 2,3,5-triphenyltetrazolium chloride (TTC) reduction as an indicator of bacterial growth. There was a good correlation with the results based on TTC reduction and the conventional method based on turbidity changes caused by bacterial growth. Furthermore, the modified method using TTC reduction is more rapid and can be read by the naked eye because of the red colour.     

Complex role of 5-HT in the regulation of TSH secretion in the male rat

The role of 5-hydroxytryptamine (5-HT) in the regulations of TSH secretion was studied in male rats using both peripheral and central administration of the drugs. Basal TSH levels were not modified by moderate doses of 5-HT (subcutaneously) or its precursors or antagonists (intraperitoneally) given 1 h before decapitation. The cold-stimulated TSH secretion was decreased by L-tryptophan (L-TRP, 400 mg/kg i.p.), quipazine (10 mg/kg i.p.) and 5-HT (1 or 5 mg/kg s.c. or i.v.) as well as by p-chlorophenylalanine (pCPA, 20 or more mg/kg i.p.) when the drugs were given 1 h before sampling. pCPA (100-400 mg/kg i.p.) was active 24-48 h after the injection but repetitive administration did not affect TSH levels. 5-HT (5 mg/kg s.c.) was effective also in pinealectomized animals. L-TRP and 5-hydroxytryptophan potentiated the TRH-stimulated TSH secretion when given 1 h before killing. 5-HT (10 microgram/rat) infused into the third ventricle enhanced the cold-stimulated TSH secretion when given 30-45 min before sampling. When injected into the medial basal hypothalamus, 50-HT (1-10 microgram/rat) had no effect on basal or stimulated TSH levels. The results suggest: (1) 5-HT does not play any role in the regulation of basal TSH secretion; (2) in the cold-stimulated TSH secretion 5-HT has a stimulatory action evidently inside the blood-brain barrier and also an inhibitory effect obviously outside this barrier.     

IFN-gamma induces a phospholipase D dependent triphasic activation of protein kinase C in endothelial cells.

The IFN-gamma linked PKC activation in endothelial cells was analysed. It was shown that IFN-gamma activates PKC in three transient and separate cycles within the first 60 minutes after IFN-gamma stimulation. Before each PKC activation there was an increase in DAG level. IP3, phosphocholine and choline productions were measured to determine the origin of DAG. Neither of the PLC products, IP3 or phosphocholine, were released after IFN-gamma stimulation. On the other hand the PLD products choline and PA were released before all the three activation cycles of PKC.     

Morphological and genetic variation of Fucus in the eastern Gulf of Bothnia,northern Baltic Sea

Fucus vesiculosus and F. radicans (Phaeophyceae) are important habitat-formers on rocky shores in the Bothnian Sea. While both species occur sympatrically along the entire western Bothnian Sea coast, F. radicans has been found only in the northern part of the eastern coast. According to previous studies, the two species can be distinguished based on morphology, F. radicans having narrower thalli and a bushier appearance. However, marine mapping in the eastern Bothnian Sea has revealed that high morphological variation in Fucus, partly caused by gradients in salinity and exposure, makes differentiation between the two species difficult. We studied morphological and genetic variation to find out whether the two Fucus species can be differentiated in the south-eastern Bothnian Sea, and if F. radicans occurs in the area. The study was carried out in six subareas including 350 km of coast, with a salinity gradient of 3.5–6.5 PSU, and varying wave exposure. We found a gradual change towards smaller and narrower thalli and a higher number of holdfasts in Fucus populations when moving northwards to lower salinities. Distinct Fucus morphs were often found within the study sites but the morphs were genetically differentiated only at one study site in the Skaftung subarea, suggesting the occurrence of both species. However, in the Vasa subarea the sample size for analysing genetic differentiation was low due to high clonality. In the Luvia subarea south of Skaftung, Fucus morphology corresponded to that of F. radicans in earlier studies but the population was genetically more similar to F. vesiculosus in the southern subareas. We conclude that by using only morphological characteristics it is not possible to differentiate between the two species in central and northern parts of the eastern Bothnian Sea. Based on genetic analyses, the southernmost known occurrence of F. radicans in the eastern Bothnian Sea is in Skaftung.     

A comparison of the host preference of monarch butterflies (Danaus plexippus) for milkweed (Asclepias syriaca) over dog-strangler vine (Vincetoxicum rossicum)

Observations in the field indicate that monarch butterflies will oviposit on dog‐strangler vine, an invasive introduced species in the same family as milkweed (Asclepias spp.), the principal larval host of monarchs. The potential impact of this behaviour depends on the strength of the preference of monarch adults to oviposit on these two hosts and the relative ability of larvae to survive on each. We determined the preference for milkweed vs. dog‐strangler vine of ovipositing adults and first instar larvae in choice and no‐choice tests. We also compared the ability of larvae to consume, develop, and survive on either host. In the presence of both hosts, adults exhibited a strong preference to oviposit on milkweed over dog‐strangler vine (mean 80.7 eggs compared to 0.4 eggs over 48 h, respectively). In the absence of milkweed, adults ceased oviposition (mean 0.9 eggs in 48 h), but resumed oviposition when the dog‐strangler vine was replaced with milkweed (mean 99.1 eggs in 48 h). Given a choice between hosts over 24 h, 92% of larvae moved to milkweed leaves and consumed 3.94 cm² of milkweed leaves compared to 2% of larvae that moved to dog‐strangler vine and consumed negligible amounts of leaf material (0.01 cm²). Without a choice, larvae on dog‐strangler vine never consumed more than mean 0.02 cm² larva^?1 in a 24‐h period, did not develop beyond the first instar, and died within 96 h. We obtained no data in support of an effect of the presence of dog‐strangler vine on monarch butterfly populations.