1-O-Alkylglycerol Ethers from the Marine Sponge Guitarra abbotti and Their Cytotoxic Activity

The cytotoxicity-bioassay-guided fractionation of the ethanol extract from the marine sponge Guitarra abbotti, whose 1-O-alkyl-sn-glycerol ethers (AGEs) have not been investigated so far, led to the isolation of a complex lipid fraction containing, along with previously known compounds, six new lipids of the AGE type. The composition of the AGE fraction as well as the structures of 6 new and 22 previously known compounds were established using ¹H and ¹³C NMR, GC/MS, and chemical conversion methods. The new AGEs were identified as: 1-O-(Z-docos-15-enyl)-sn-glycerol (1), 1-O-(Z-docos-17-enyl)-sn-glycerol (2), 1-O-(Z-tricos-15-enyl)-sn-glycerol (3), 1-O-(Z-tricos-16-enyl)-sn-glycerol (4), 1-O-(Z-tricos-17-enyl)-sn-glycerol (5), and 1-O-(Z-tetracos-15-enyl)-sn-glycerol (6). The isolated AGEs show weak cytotoxic activity in THP-1, HL-60, HeLa, DLD-1, SNU C4, SK-MEL-28, and MDA-MB-231 human cancer cells. A further cytotoxicity analysis in JB6 P⁺ Cl41 cells bearing mutated MAP kinase genes revealed that ERK2 and JNK1 play a cytoprotective role in the cellular response to the AGE-induced cytotoxic effects.     

Evaluation of non-chemical seed treatment methods for the control of Alternaria dauci and A. radicina on carrot seeds

The current study was initiated to evaluate the efficacy of physical methods (hot water, aerated steam, electron treatment) and agents of natural origin (resistance inducers, plant derived products, micro-organisms) as seed treatments of carrots for control of Alternaria dauci and A. radicina. Control of both Alternaria species by seed treatment with the resistance inducers was generally poor. Results were also not satisfactory with most of the formulated commercial micro-organism preparations. Based on the average of five field trials, one of these, BA 2552 (Pseudomonas chlororaphis), provided a low but significant increase in plant stand. Among the experimental micro-organisms, the best results were obtained with Pseudomonas sp. strain MF 416 and Clonostachys rosea strain IK726. A similar level of efficacy was provided by seed treatment with an emulsion (1%) of thyme oil in water. Good and consistent control was generally achieved with the physical methods aerated steam, hot water and electron treatment. Aerated steam treatment was, apart from the thiram-containing chemical standard, the best single treatment, and its performance may at least partially be due to extensive pre-testing, resulting in dosages optimally adapted to the respective seed lot. In some of the experiments the effect of the hot water treatment, which was tested at a fixed, not specifically adapted dosage, was significantly improved when combined with a Pseudomonas sp. MF 416 or C. rosea IK726 treatment. The results are discussed in relation to the outcome of experiments in which the same seed treatment methods and agents were tested in other seed-borne vegetable pathosystems.     

The use of small (2.7 mm) screws for arthroscopically guided repair of carpal chip fractures

Reasons for performing the study: Removal of large chip fractures of the carpal bones and the osteochondral deficits that result, have been associated with a worse prognosis than removal of small fragments in similar locations. Hypothesis: Reducing the articular defects by repair of large osteochondral fragments may have advantages over removal. Methods: Horses with osteochondral chip fractures that were of sufficient size and infrastructure to be repaired with small (2.7 mm diameter) AO/ASIF cortex screws were identified and repair effected by arthroscopically guided internal fixation. Results: Thirty‐three horses underwent surgery to repair 35 fractures of the dorsodistal radial carpal bone (n = 25), the dorsal margin of the radial facet of the third carpal bone (n = 9) and the intermediate facet of the distal radius (n = 1). There were no surgical complications and fractures healed satisfactorily in 26 of 28 horses and 23 horses returned to racing performance. Conclusion: Arthroscopically guided repair of carpal chip fractures with small diameter cortex screws is technically feasible and experiences with 33 cases suggest that this may have advantages over fragment removal in managing such cases. Potential relevance: Surgeons treating horses with large chip fractures of the carpal bones should consider arthroscopically guided internal fixation as an alternative to removal.     

Type 2 diabetes mellitus with pancreatic β cell dysfunction in 3 horses confirmed with minimal model analysis

Reasons for performing study: Type 2 diabetes mellitus (T2DM) is diagnosed rarely in equine practice although it may be under‐recognised. A greater awareness of the condition and therapeutic considerations would be to the benefit of such cases presenting in practice. More investigation into the pharmacological management of these cases is needed. Objectives: Three cases of diabetes mellitus were investigated using a specific test for insulin sensitivity and pancreatic β cell function in order to define accurately and characterise the existence of T2DM in all 3 subjects. Methods: The insulin‐modified frequently sampled i.v. glucose tolerance test was performed in each case and the data so obtained were subject to minimal model analysis of insulin‐glucose dynamics. Cases were then monitored following treatment using a combination of dietary modification, metformin, glibenclamide and pergolide. Results: Marked insulin resistance was identified in each case and, furthermore, severe pancreatic β cell dysfunction was present therefore classifying each case as end stage T2DM. Treatment was nevertheless associated with restoration of normoglycaemia in all cases. Conclusions: T2DM in horses may be more common than generally considered. In some cases individuals may respond to therapy aimed at restoring insulin sensitivity and pancreatic function. Drugs used in other species for the treatment of T2DM have not yet been adequately tested in horses. Potential relevance: T2DM should be considered as an important differential diagnosis in mature to elderly horses and ponies suffering from weight loss, polydipsia and polyuria. Clinicians should be encouraged to offer treatment and management advice when such cases are encountered.     

Pharmacodynamics of amoxicillin against Mannheimia haemolytica and Pasteurella multocida and pharmacokinetic/pharmacodynamic (PK/PD) correlation in sheep

Silicone-made tissue cages were implanted in sheep. Blood serum (SBS) and tissue cage fluid (TCF) samples were collected after amoxicillin intravenous and intramuscular administrations, at the dose of 15 mg/kg. Amoxicillin pharmacodynamics were studied in an artificial culture medium, SBS and TCF with use of a Mannheimia haemolytica and a Pasteurella multocida strain. A concentration-independent antimicrobial activity of amoxicillin was confirmed for levels higher than 0.79–1.75 × MIC. This result favored the use of the percentage of the 24 h dosing interval during which drug levels remain above MIC as the appropriate pharmacokinetic/pharmacodynamic index. The subsequent correlation revealed that intravenous administration could be considered effective against “deep” infections caused by bacteria with MICs < 1 μg/mL or “shallow” infections caused by bacteria with MICs < 0.1 μg/mL. Intramuscular administration could be safely considered effective against both “deep” and “shallow” infections when the MICs of the targeted pathogens are lower than 1 μg/mL.     

Effect of endobronchial challenge with Actinobacillus pleuropneumoniae serotype 10 of pigs vaccinated with bacterins consisting of A. pleuropneumoniae serotype 10 grown under NAD-rich and NAD-restricted conditions

The efficacy of two bacterins containing an Actinobacillus pleuropneumoniae serotype 10 strain was evaluated. The bacterial cells constituting bacterin 1 and 2 were grown under nicotinamide adenine dinucleotide (NAD)-rich (low-adherence capacity to alveolar epithelial cell cultures) and NAD-restricted (high-adherence capacity to alveolar epithelial cell cultures) conditions, respectively. Ten pigs were vaccinated twice with the bacterin 1 and nine pigs with the bacterin 2. Ten control animals were injected twice with a saline solution. Three weeks after the second vaccination, all pigs were endobronchially inoculated with 106.5 colony-forming units (CFU) of an A. pleuropneumoniae serotype 10 strain. In the bacterin 1 and 2 group, three and two pigs died after inoculation, respectively. Only two pigs of the control group survived challenge. Surviving pigs were killed at 7 days after challenge. The percentage of pigs with severe lung lesions (> 10% of the lung affected) was 100% in the control group, 70% in the bacterin 1 group and 22% in the bacterin 2 group. Actinobacillus pleuropneumoniae was isolated from the lungs of all animals. The mean bacterial titres of the caudal lung lobes were 7.0 x 10(6) CFU/g in the control group, 6.3 x 10(5) CFU/g in the bacterin 1 group and 1.3 x 10(6) CFU/g in the bacterin 2 group. It was concluded that both bacterins induced partial protection against severe challenge. Furthermore, there are indications that the bacterin 2, containing A. pleuropneumoniae bacteria grown under conditions resulting in high in vitro adhesin, induced better protection than the bacterin 1.     

A Comparison of Simple Continuous Versus Simple Interrupted Suture Patterns for Tracheal Anastomosis After Large-Segment Tracheal Resection in Dogs

Two suture techniques for tracheal anastomosis after large-segment tracheal resection were compared. Eight cartilages were resected from the trachea of each of 12 dogs; anastomoses with 4-0 polydioxanone suture were created using a simple continuous suture technique in six dogs and a simple interrupted suture technique in six dogs. Surgical time was shorter but apposition of tracheal segments at the time of surgery was less precise with the simple continuous suture technique. The dogs were evaluated for 150 days after surgery. Clinical abnormalities after tracheal resection and anastomosis were not observed. Percent dorsoventral luminal stenosis was calculated by measuring the tracheal lumen diameter on lateral cervical radiographs. Percent luminal stenosis was calculated planimetrically using a computerized digitizing tablet. Anastomotic stenosis was mild in all dogs; however, the mean percent luminal stenosis determined planimetrically was significantly greater for dogs that had the simple continuous suture technique. Planimetric measurements of cross-sectional area made before and after formalin fixation were not significantly different. Radiographic determination of percent dorsoventral luminal stenosis was a poor predictor of diminution of cross-sectional area determined planimetrically.     

Arachidonic acid immunoregulation in lambs persistently infected with border disease virus

To evaluate arachidonic acid-related immunoregulatory mechanisms during long-term persistent pestivirus infection, we measured plasma contents of leukotriene C4 (LTC4), prostaglandin D2 (PGD2) and their plasma fatty acid (FA) precursor, arachidonic acid (AA), in six lambs with congenital border disease (BD). Significantly elevated average plasma LTC4 during the first half year of life was associated with increased PDG2 when compared to uninfected control lambs. Significantly elevated total plasma esterified AA stores suggest an effective BDV-mediated prostenoid immunostimulation. However, at 1 yr old, prostenoid secretion had fallen to normal (LTC4) or below normal (PGD2) levels. In contrast, there remained significantly elevated plasma esterified AA, present as available substrate for formation of these anti-viral immunoregulatory agents. These results suggested that preventing mobilization of AA from lipid stores for effective immune responses may be a viral strategy of BD virus that is associated with long term border disease effects.