β-Carotene from the Alga Dunaliella bardawil Decreases Gene Expression of Adipose Tissue Macrophage Recruitment Markers and Plasma Lipid Concentrations in Mice Fed a High-Fat Diet

Vitamin A and provitamin A carotenoids are involved in the regulation of adipose tissue metabolism and inflammation. We examined the effect of dietary supplementation using all-trans and 9-cis β-carotene-rich Dunaliella bardawil alga as the sole source of vitamin A on obesity-associated comorbidities and adipose tissue dysfunction in a diet-induced obesity mouse model. Three-week-old male mice (C57BL/6) were randomly allocated into two groups and fed a high-fat, vitamin A-deficient diet supplemented with either vitamin A (HFD) or β-carotene (BC) (HFD-BC). Vitamin A levels in the liver, WATs, and BAT of the HFD-BC group were 1.5–2.4-fold higher than of the HFD group. BC concentrations were 5–6-fold greater in BAT compared to WAT in the HFD-BC group. The eWAT mRNA levels of the Mcp-1 and Cd68 were 1.6- and 2.1-fold lower, respectively, and the plasma cholesterol and triglyceride concentrations were 30% and 28% lower in the HFD-BC group compared with the HFD group. Dietary BC can be the exclusive vitamin A source in mice fed a high-fat diet, as shown by the vitamin A concentration in the plasma and tissues. Feeding BC rather than vitamin A reduces adipose tissue macrophage recruitment markers and plasma lipid concentrations.     

Upfront denileukin diftitox as in vivo regulatory T-cell depletion in order to enhance vaccination effects in a canine allogeneic hematopoietic stem cell transplantation model

Denileukin Diftitox (ONTAK^®, DAB₃₈₉ IL-2) is a recombinant DNA-derived fusion protein depleting cells that express high-affinity IL-2 receptor. Important cell targets are CD4⁺CD25⁺Foxp3⁺ regulatory T cells (T_reg). Elimination of immunosuppressive T_reg by Denileukin Diftitox may provide a way to modulate immune tolerance following stem cell transplantation. Here, we combined T_reg depletion with a vaccination approach to induce donor-specific immune reactions. To investigate this approach we chose the mixed chimerism canine stem cell transplantation model which represents a high state of tolerance between two hematopoietic systems. The aim was therefore to induce a graft versus hematopoiesis effect thereby converting mixed to full donor chimerism. Dog leukocyte antigen identical siblings that had developed a stable mixed chimerism after non-myeloablative stem cell transplantation received a single dose of Denileukin Diftitox (18 μg/kg, i.v.) followed by several cell-lysate vaccinations. Host peripheral blood mononuclear cell lysates combined with CpG-ODN, and Montanide^® ISA 51 were locally applied. In vitro studies demonstrated that canine T_reg are a target of Denileukin Diftitox. The suppression of T-cell proliferation by T_reg was abolished by addition of Denileukin Diftitox (10 nM). An increase of proliferation of median 300% (range: 200%–425%) was observed. No change in donor chimerism was observed after administration of Denileukin Diftitox and vaccination. This study highlights that application of Denileukin Diftitox resulted in a depletion of T_reg followed by an increase of immune response in vitro. This effect could not be confirmed in vivo even if the immune system was stimulated by vaccinations.     

Presence of Muscarinic Acetylcholine Receptors in the Cattle Tick Boophilus microplus and in Epithelial Tissue Culture Cells of Chironomus tentans

A muscarinic acetylcholine receptor (mAChR) has been demonstrated and partially characterized in larvae of the cattle tick Boophilus microplus. Its properties are compared with mAChR from an epithelial cell line from the dipteran insect Chironomus tentans. Competition studies with cholinergic ligands of different specificity revealed the muscarinic nature of the cholinergic receptors investigated in both species. In homogenates from tick larvae, specific binding sites for [³H]quinuclidinyl benzilate (QNB) with high affinity (1·2±(0·13) nM ; B_max 22·5 pmol mg protein⁻¹) were detected that do not bind nicotinic compounds specifically. The estimated IC₅₀ values for nicotine, imidacloprid and α-bungarotoxin were all in the mM range. Additionally, with tick larvae, high-affinity nicotinic binding sites were detected with [³H]nicotine which could be displaced by high concentrations of imidacloprid or QNB. The estimated IC₅₀ values for nicotine, α-bungarotoxin, imidacloprid and QNB were 43(±8) nM , 0·8(±0·2) μM , 2·8(±0·6) μM and 78(±1·9) μM , respectively. With homogenates of the non-neuronal insect cell line from C. tentans, only high-affinity binding sites for [³H]QNB were found. Muscarinic antagonists selectively displaced [³H]quinuclidinyl benzilate (QNB) binding to tick larvae homogenates. The mAChR of B. microplus preferred pirenzepine (IC₅₀ 2·13(±1·02) μM ) among different subtype-specific mAChR antagonists (4-DAMP had IC₅₀ 49·9(±9·13) μM and methoctramine had IC₅₀ 121(±14·2) μM ) indicating a type of binding site similar to the vertebrate M1 mAChR subtype. The tick muscarinic receptor seems to be a G-protein-coupled receptor, as concluded from the 4·8-fold reduction in receptor affinity for binding of the muscarinic agonist oxotremorine M upon treatment with the non-hydrolysable GTP-analogue γ-S-GTP. Binding data for the agonists oxotremorine M (IC₅₀ 71·3(±19·6) μM ) and carbachol (IC₅₀ 253(±87·1) μM ) parallel the biological efficacy of these compounds, in that, while oxotremorine M showed some activity against ticks, carbachol was ineffective.     

Laryngeal mask airway insertion requires less propofol than endotracheal intubation in dogs

OBJECTIVE: To compare the doses of propofol required for insertion of the laryngeal mask airway (LMA) with those for endotracheal intubation in sedated dogs. STUDY DESIGN: Randomized prospective clinical study. Animals Sixty healthy dogs aged 0.33-8.5 (3.0 +/- 2.3, mean +/- SD) years, weighing 2.2-59.0 (23.4 +/- 13.6, mean +/- SD) kg, presented for elective surgery requiring inhalation anaesthesia. METHODS: Animals were randomly assigned to receive either a LMA or an endotracheal tube. Pre-anaesthetic medication was intravenous (IV) glycopyrrolate (0.01 mg kg(-1)) medetomidine (10 microg kg(-1)) and butorphanol (0.2 mg kg(-1)). Repeated IV propofol injections (1 mg kg(-1) in 30 seconds) were given until LMA insertion or endotracheal intubation was achieved, when the presence or absence of laryngospasm, the respiratory rate (fr) and the total dose of propofol used were recorded. RESULTS: The total propofol dose (mean +/- SD) required for LMA insertion (0.53 +/- 0.51 mg kg(-1)) was significantly lower than for endotracheal intubation (1.43 +/- 0.57 mg kg(-1)). The LMA could be inserted without propofol in 47% of dogs; the remainder needed a single 1 mg kg(-1) bolus (n = 30). Endotracheal intubation was possible without propofol in 3.3% of the dogs, 47% needed one bolus and 50% required two injections (n = 30). The f(r) (mean +/- SD) was 18 +/- 6 and 15 +/- 7 minute(-1) after LMA insertion and intubation, respectively. CONCLUSION AND CLINICAL RELEVANCE: Laryngeal mask airway insertion requires less propofol than endotracheal intubation in sedated dogs therefore propofol-induced cardiorespiratory depression is likely to be less severe. The LMA is well tolerated and offers a less invasive means of securing the upper airway.     

Laser ablation-combustion-GC-IRMS--a new method for online analysis of intra-annual variation of delta13C in tree rings

We present a new, rapid method for high-resolution online determination of delta13C in tree rings, combining laser ablation (LA), combustion (C), gas chromatography (GC) and isotope ratio mass spectrometry (IRMS) (LA-C-GC-IRMS). Sample material was extracted every 6 min with a UV-laser from a tree core, leaving 40-microm-wide holes. Ablated wood dust was combusted to CO2 at 700 degrees C, separated from other gases on a GC column and injected into an isotope ratio mass spectrometer after removal of water vapor. The measurements were calibrated against an internal and an external standard. The tree core remained intact and could be used for subsequent dendrochronological and dendrochemical analyses. Cores from two Scots pine trees (Pinus sylvestris spp. sibirica Lebed.) from central Siberia were sampled. Inter- and intra-annual patterns of delta13C in whole-wood and lignin-extracted cores were indistinguishable apart from a constant offset, suggesting that lignin extraction is unnecessary for our method. Comparison with the conventional method (microtome slicing, elemental analysis and IRMS) indicated high accuracy of the LA-C-GC-IRMS measurements. Patterns of delta13C along three parallel ablation lines on the same core showed high congruence. A conservative estimate of the precision was +/- 0.24 per thousand. Isotopic patterns of the two Scots pine trees were broadly similar, indicating a signal related to the forest stand's climate history. The maximum variation in delta13C over 22 years was about 5 per thousand, ranging from -27 to -22.3 per thousand. The most obvious pattern was a sharp decline in delta13C during latewood formation and a rapid increase with spring early growth. We conclude that the LA-C-GC-IRMS method will be useful in elucidating short-term climate effects on the delta13C signal in tree rings.     

Case–control risk factor study of methicillin-resistant Staphylococcus pseudintermedius (MRSP) infection in dogs and cats in Germany

Methicillin-resistant Staphylococcus pseudintermedius (MRSP) has emerged as a highly drug-resistant small animal veterinary pathogen. Although often isolated from outpatients in veterinary clinics, there is concern that MRSP follows a veterinary-hospital-associated epidemiology. This study's objective was to identify risk factors for MRSP infections in dogs and cats in Germany. Clinical isolates of MRSP cases (n = 150) and methicillin-susceptible S. pseudintermedius (MSSP) controls (n = 133) and their corresponding host signalment and medical data covering the six months prior to staphylococcal isolation were analysed by multivariable logistic regression. The identity of all MRSP isolates was confirmed through demonstration of S. intermedius-group specific nuc and mecA. In the final model, cats (compared to dogs, OR 18.5, 95% CI 1.8–188.0, P = 0.01), animals that had been hospitalised (OR 104.4, 95% CI 21.3–511.6, P < 0.001), or visited veterinary clinics more frequently (>10 visits OR 7.3, 95% CI 1.0–52.6, P = 0.049) and those that had received topical ear medication (OR 5.1, 95% CI 1.8–14.9, P = 0.003) or glucocorticoids (OR 22.5, 95% CI 7.0–72.6, P < 0.001) were at higher risk of MRSP infection, whereas S. pseudintermedius isolates from ears were more likely to belong to the MSSP-group (OR 0.09, 95% CI 0.03–0.34, P < 0.001). These results indicate an association of MRSP infection with veterinary clinic/hospital settings and possibly with chronic skin disease. There was an unexpected lack of association between MRSP and antimicrobial therapy; this requires further investigation but may indicate that MRSP is well adapted to canine skin with little need for selective pressure.     

Modification of Marine Natural Product Ningalin B and SAR Study Lead to Potent P-Glycoprotein Inhibitors

In this study, new marine ningalin B analogues containing a piperazine or a benzoloxy group at ring C have been synthesized and evaluated on their P-gp modulating activity in human breast cancer and leukemia cell lines. Their structure-activity relationship was preliminarily studied. Compounds 19 and 20 are potent P-gp inhibitors. These two synthetic analogues of permethyl ningalin B may be potentially used as effective modulators of P-gp-mediated drug resistance in cancer cells.     

Breeding performance of the second generation in some bivoltine populations of <Emphasis Type="Italic">Ips typographus</Emphasis> (Coleoptera Curculionidae) in the south-eastern Alps

The spruce bark beetle, Ips typographus, is one of the most extensively studied European forest pests. Gaps exist in the knowledge about second generation breeding performance in bivoltine populations. In this study, the breeding performance of the second generation was evaluated in three bivoltine populations of I. typographus in the SE Alps. Length of the maternal galleries (from 40.5 to 44.8 mm), population growth rate (PGR; from 0.7 to 3.6), and emerged adults per m² (from 669 to 1,570 insects/m²) varied among populations and were negatively correlated with bark colonisation density. Pheromone traps set up in the three investigated forests differed in the number of trapped beetles, with mean captures ranging between 5,310 and 19,850 insects per trap. The populations giving the highest captures in the traps showed the lowest bark colonisation density (248 vs. 489 maternal galleries per m²) and the best breeding performance. The populations of parasitoids and predators corresponded to just 1–9 and 2–10% of the emerging I. typographus adults, respectively, and phloem temperature never reached thresholds lethal to I. typographus. Interspecific competition was negligible, whereas intraspecific competition was found to be the main factor affecting the breeding performance of the second generation, although with different intensity according to the colonisation density. It is hypothesised that competition with the first generation and spring precipitation influence the number of suitable hosts available to the second generation.