Bile acids stimulate PKCalpha autophosphorylation and activation: role in the attenuation of prostaglandin E1-induced cAMP production in human dermal fibroblasts

The aim was to identify the specific PKC isoform(s) and their mechanism of activation responsible for the modulation of cAMP production by bile acids in human dermal fibroblasts. Stimulation of fibroblasts with 25-100 microM of chenodeoxycholic acid (CDCA) and ursodeoxycholic acid (UDCA) led to YFP-PKCalpha and YFP-PKCdelta translocation in 30-60 min followed by a transient 24- to 48-h downregulation of the total PKCalpha, PKCdelta, and PKCepsilon protein expression by 30-50%, without affecting that of PKCzeta. Increased plasma membrane translocation of PKCalpha was associated with an increased PKCalpha phosphorylation, whereas increased PKCdelta translocation to the perinuclear domain was associated with an increased accumulation of phospho-PKCdelta Thr505 and Tyr311 in the nucleus. The PKCalpha specificity on the attenuation of cAMP production by CDCA was demonstrated with PKC downregulation or inhibition, as well as PKC isoform dominant-negative mutants. Under these same conditions, neither phosphatidylinositol 3-kinase, p38 MAP kinase, p42/44 MAP kinase, nor PKA inhibitors had any significant effect on the CDCA-induced cAMP production attenuation. CDCA concentrations as low as 10 microM stimulated PKCalpha autophosphorylation in vitro. This bile acid effect required phosphatidylserine and was completely abolished by the presence of G?6976. CDCA at concentrations less than 50 microM enhanced the PKCalpha activation induced by PMA, whereas greater CDCA concentrations reduced the PMA-induced PKCalpha activation. CDCA alone did not affect PKCalpha activity in vitro. In conclusion, although CDCA and UDCA activate different PKC isoforms, PKCalpha plays a major role in the bile acid-induced inhibition of cAMP synthesis in fibroblasts. This study emphasizes potential consequences of increased systemic bile acid concentrations and cellular bile acid accumulation in extrahepatic tissues during cholestatic liver diseases.     

Duplex sonography of arteriovenous fistula in chronic hemodialysis patients

Duplex sonography was used to assess functional features of arteriovenous fistula (AVF) for hemodialysis (HD). Internal diameter (ID), resistance index (RI) and blood flow (BF) velocity in feeding artery and in vein ofAVF, and venous BF volume were analyzed with purpose to determine the normal values. Presumed normal BF velocities are those of clinically well functioning shunts, allowing BF through HD lines of minimally 250 ml/min. Study included 66 nondiabetic HDpatients (30 women, 36 men), mean age 52-13 years, treated by HD for median 61 (4-252) months. Measurements in 47patients with clinically well functioning AVF were as followed: mean arterial ID 5.2 +/- 1.4 mm, median arterial RI 0.3 (0.3-0.9), median arterial BF velocity 1.5 (0.6-3.6) m/s, mean venous ID 7.6 +/- 2.2 mm, median venous RI 0.3 (0.3-0.9), mean venous BF velocity 1.6 +/- 0.7 m/s, and median venous BF volume 530 (120-1890) ml/min. Patients with poor functioning AVF had significantly less arterial ID, higher arterial RI, less venous ID, less venous BF velocity and volume. Duplex sonography findings obtained for clinically estimated well functioning shunt should be considered as normal Doppler values. Blood vessels' morphologic features depend upon age, and older patients have more pronounced changes.     

Distribution of anuran amphibians in massively altered landscapes in south-eastern Australia: effects of climate change in an aridifying region

Aim We explored whether the anuran amphibian faunas differed among landscapes that are relatively intact (largely covered in forests and woodlands) and others that are completely converted to agriculture. We also used historical data sets to assess the current condition of the anuran fauna in a region predicted to experience, and experiencing, severe drying and warming. Location Five pairs of landscapes (each of c. 20 km²) – one in each pair being almost completely wooded and the other cleared for agriculture – across a 30,000 km² region of northern Victoria, Australia. Methods Sites were repeatedly surveyed in the austral winter–spring breeding seasons of 2006 and 2007, with records collected of numbers of calling males and the presence of egg masses and tadpoles. We characterized the sites using static (e.g. dimensions, surrounding physiognomic characteristics such as tree cover) and labile (e.g. pH, dissolved oxygen) variables. Data were analysed using hierarchical Bayesian models. Results For calling males, landscape type did not affect densities or species richness measures. The availability of a grassy verge around water bodies was an important predictor for most species, but other labile and static variables seemed not to be important. Fewer than half of the species historically known from the region were recorded. There were no important predictors of occurrence of egg masses or tadpoles. Reproduction effectively may have failed over the period, with fewer than one in four sites showing evidence of egg masses or tadpoles. Main conclusions The proportion of sites at which some well‐studied species (e.g. Crinia signifera, Limnodynastes dumerilii) were recorded has dropped substantially since the 1970s, as have average densities of calling males of Crinia spp. The remnant anuran fauna appears to be dominated by resilient and hardy species with low current diversity. The on‐going (12+ years) drought in these landscapes suggests a bleak long‐term prognosis for the few remaining species of anuran amphibians.     

The Benefits of Ambiol® in Promoting Germination, Growth, and Drought Tolerance can be Passed on to Next-Generation Tomato Seedlings

Seed preconditioning with Ambiol^® has been shown to improve germination, growth, and drought tolerance in seedlings of many species. Attempts to understand the mode of action of Ambiol have found that seed preconditioning triggers several new proteins, which suggests that Ambiol-induced benefits may persist beyond seedlings and, perhaps, into the next generation. Seeds were preconditioned with 0 and 10 mg l^?1 Ambiol to determine effects on germination, seedling growth, and yield of parent tomato plants. Seeds were collected from plants in each treatment and then sown to determine effects on germination and seedling growth in the next generation. Key parameters such as percent germination, leaf area, shoot mass, root mass, and photosynthesis were significantly improved in parents and in progeny. In addition, there was a 141% increase in tomato yield in preconditioned parents. It was concluded that Ambiol-induced benefits continue throughout plant development and into the next generation, potentially having significant horticultural and economic ramifications.     

Natural infection of Cryptosporidium muris (Apicomplexa: Cryptosporiidae) in Siberian chipmunks

Coprologic examination of nine Siberian chipmunks (Eutamias sibiricus) imported from Southeast Asia revealed infection with Cryptosporidium sp. Experimental inoculation of BALB/c mice proved their susceptibility to the infection. Infected mice shed oocysts 14-35 days postinfection. Oocyst morphology was similar to that reported for C. muris in previous studies, oocysts were 8.1 (7.0-9.0) x 5.9 (5.0-6.5) microns. Clinical signs were absent in naturally infected chipmunks and experimental mice. Histologic examinations of mice revealed numerous developmental stages of C. muris in the glandular stomach. Analysis of partial small subunit rRNA gene sequences confirmed identity of these isolates as C. muris. Our results represent the first report of C. muris in members of the family Sciuridae.     

Evidence for Neuronal Origin and Metabotropic Receptor-Mediated Regulation of Extracellular Glutamate and Aspartate in Rat Striatum In Vivo Following Electrical Stimulation of the Prefrontal Cortex

Abstract: Extracellular levels of glutamate (Glu) and aspartate (Asp) were measured at 5-s intervals in the striatum of chloral hydrate-anesthetized rats by using microdialysis coupled to an automated assay system based on capillary electrophoresis with laser-induced fluorescence. Application of a single 10-s train of depolarizing pulses to the prefrontal cortex caused a rapid increase in Glu and Asp concentrations (200–300% of basal value), which returned to basal level within 60 s. The stimulated rise in Glu and Asp concentrations was blocked completely by 2 µ M tetrodotoxin or depletion of extracellular Ca²⁺, suggesting a neuronal origin of the Glu and Asp. Infusion of the Glu transport inhibitor l - trans -pyrrolidine-2,4-dicarboxylic acid (200 µ M ) increased resting Glu and Asp levels by 300–500% without altering electrically stimulated changes in Glu and Asp concentration. Stimulated Glu and Asp concentration changes were suppressed by 91 and 73%, respectively, by the metabotropic Glu receptor agonist (1 S ,3 R )-1-aminocyclopentane- trans -1,3-dicarboxylate (200 µ M ). This effect was blocked by the metabotropic Glu receptor antagonist ( RS )-α-methylcarboxyphenylglycine (MCPG; 200 µ M ). MCPG alone produced no effect on electrically stimulated changes in Glu and Asp levels; however, in the presence of l - trans -pyrrolidine-2,4-dicarboxylic acid, MCPG produced a five- to sixfold increase in stimulated overflow. Based on these results, it is concluded that release of Glu and Asp from corticostriatal neurons can be inhibited by activation of metabotropic Glu autoreceptors, which may be an important determinant of excitatory transmission at striatal synapses.     

Early growth invigoration of jack pine seedlings by natural plant growth regulators

 Survival of conifer transplants is often poor on exposed planting sites in the boreal forest. More than one-third of all conifers do not become established. To enhance the competitive ability of jack pine seedlings, seeds were treated with natural plant growth regulators (PGRs; viz., homobrassinolide, salicylic acid, and two polyamines, spermine and spermidine) and growth promotion was studied for 16 days. Homobrassinolide (5 ng l^–1), salicylic acid (100 μg l^–1) and spermine (10 μg l^–1) enhanced elongation growth and elongation rate of whole plant. Homobrassinolide (5 ng l^–1) and salicylic acid (100 μg l^–1) stimulated root elongation by 38% and 10% respectively while spermine (1000 μg l^–1) increased needle growth by 14%. Homobrassinolide stimulated dry weight growth and growth rate. Homobrassinolide recorded over 20% increase in dry matter production, apportioned equally to root and needles, whereas spermine enhanced total dry matter production by almost 10%, mostly by increasing needle weight. Homobrassinolide facilitated nearly 19% increase in growth rate while spermine recorded only a 7% growth promotion. Spermidine inhibited both elongation and dry weight growth at all concentrations. Growth promotion by homobrassinolide, salicylic acid and spermine may be through an acceleration of processes connected to cell elongation, cell division and C allocation and these PGRs showed most promise for the early invigoration and improvement of jack pine seedling growth. Received: 27 August 1997 / Accepted: 8 January 1998     

Decline and potential recovery of Yellow-footed Antechinus in parts of south-eastern Australia: A perspective with implications for management

Summary   The extent and intensity of European-induced changes to ecosystems in south-eastern Australia mean that remaining habitats, despite being degraded, are of high conservation value. Given the extinction of several species of native mammals in the last 160 years in the area, and conservation concerns regarding others, it is important to provide conservation managers with sufficient information to prevent further extinctions and maintain evolutionary potential of the species. A native, carnivorous marsupial, the Yellow-footed Antechinus ( Antechinus flavipes ) exists within these massively altered landscapes. We present conceptual models, derived from the literature, of persistence of populations of Yellow-footed Antechinus both before European colonization and in the 21st century. We conclude that preservation of large trees, restoration of fallen-timber volumes, spring flooding of floodplains and presence of vegetation corridors between forests should be undertaken to prevent local extinctions of Yellow-footed Antechinus. From historic and current gene flow, we identify remnant woodland and forest groupings that we consider should be managed as coherent units.     

Low stimulated IL-4 secretion in PBMC from patients with chronic idiopathic urticaria

An imbalance between the Th1 and Th2 arms of the cellular immune system has been reported in several autoimmune diseases but not in chronic idiopathic urticaria (CIU). Peak, total secretion and secretory pattern of the Th1 cytokines (IFNgamma and IL-2) and Th2 cytokines (IL-4 and IL-10) were determined in resting and stimulated peripheral blood mononuclear cells (PBMC) from nineteen CIU patients, six acute urticaria patients and twelve controls. Stimulated IL-4 secretion was significantly reduced in CIU patients as indicated by their five- and three-fold lower peak levels and total IL-4 secretion, respectively. The IL-4 secretory pattern overtime was also markedly different in patients and controls. The late secretion of IFNgamma at 144 h was also reduced in CIU patients. These aberrations were not detectable in AU patients. Secretion of IL-2 was lower in CIU and AU patients as compared to controls while IL-10 secretion was comparable in the three groups. Our data demonstrate for the first time a predominantly reduced IL-4 secretion in CIU patients. This is associated with reduced secretion of both IL-2 and IFNgamma. These findings indicate a generalized down-regulation of both Th1 and Th2 cytokines' secretion in CIU.     

Effects of rHuEPO treatment on red blood cell osmotic resistance

Red blood cell osmotic resistance (RBCOR) is defined as resistance to osmotic changes in cell integrity after their exposure to hypotonic saline solution. The investigation examined the effect of rHuEPO on RBCOR in hemodialysed patients. The study included 58 patients aged 49 +/- 14 years, treated by hemodialysis for 59 +/- 43 months on average. Half of the patients received rHuEPO for anemia correction. RBCOR was determined in all patients as 3 values: hemolysis start point (HSP), hemolysis end point (HEP) and middle osmotic resistance (MOR). The patients underwent laboratory checkup for parameters characteristically changed in the uremic syndrome. In the control group of healthy subjects (n = 16) RBCOR was only determined. No differences were found in the average values of HSP, HEP and MOR between the rHuEPO treated group of patinets and the untreated group. Compared to healthy individuals, the hemodialysed patients displayed significantly higher values of HSP, HEP and MOR. The only one significant correlation of RBCOR and routine laboratory features was found between MOR and predialytic serum concentrations of calcium (r = 0.28, p < 0.05) and hydrogen ions (r = 0.37, p < 0.05). Our results suggest that the administration of rHuEPO does not affect RBCOR in hemodialysed patients, that RBCOR is not always reduced in this population and that it correlates with a small number of laboratory parameters characteristic for the uremic syndrome.