Fucose-Rich Sulfated Polysaccharides from Two Vietnamese Sea Cucumbers Bohadschia argus and Holothuria (Theelothuria) spinifera: Structures and Anticoagulant Activity

Fucosylated chondroitin sulfates (FCSs) FCS-BA and FCS-HS, as well as fucan sulfates (FSs) FS-BA-AT and FS-HS-AT were isolated from the sea cucumbers Bohadschia argus and Holothuria (Theelothuria) spinifera, respectively. Purification of the polysaccharides was carried out by anion-exchange chromatography on DEAE-Sephacel column. Structural characterization of polysaccharides was performed in terms of monosaccharide and sulfate content, as well as using a series of non-destructive NMR spectroscopic methods. Both FCSs were shown to contain a chondroitin core [→3)-β-d-GalNAc-(1→4)-β-d-GlcA-(1→]_n bearing sulfated fucosyl branches at O-3 of every GlcA residue in the chain. These fucosyl residues were different in pattern of sulfation: FCS-BA contained Fuc2S4S, Fuc3S4S and Fuc4S at a ratio of 1:8:2, while FCS-HS contained these residues at a ratio of 2:2:1. Polysaccharides differed also in content of GalNAc4S6S and GalNAc4S units, the ratios being 14:1 for FCS-BA and 4:1 for FCS-HS. Both FCSs demonstrated significant anticoagulant activity in clotting time assay and potentiated inhibition of thrombin, but not of factor Xa. FS-BA-AT was shown to be a regular linear polymer of 4-linked α-L-fucopyranose 3-sulfate, the structure being confirmed by NMR spectra of desulfated polysaccharide. In spite of considerable sulfate content, FS-BA-AT was practically devoid of anticoagulant activity. FS-HS-AT cannot be purified completely from contamination of some FCS. Its structure was tentatively represented as a mixture of chains identical with FS-BA-AT and other chains built up of randomly sulfated alternating 4- and 3-linked α-L-fucopyranose residues.     

Removal of flowers or inflorescences affects ‘Barnea’ olive fruitlet post-anthesis abscission

A typical olive (Olea europaea L.) inflorescence consists of about 20 flowers. However, in many cultivars, only one fruit develops. This is due to massive abscission of flowers and fruitlets, which occurs during the first month after anthesis. In this study, we used the olive cultivar 'Barnea' to characterize the abscission mechanism and to try to increase fruit set by increasing the number of developed fruit per inflorescence. Removing the lateral flowers 3 weeks before anthesis increased fruit set by more than 50%. Removing all inflorescences but one from a branch increased the number of developed fruits from 0.93 to 2.8 during 2017 and from 0.91 to 3.34 fruits per inflorescence in 2018. Sugar quantification in the pistil revealed that starch level is high on the day of anthesis and low 25 days later in abscised as well as in developed fruit. Soluble carbohydrates are low on the day of anthesis, low in abscised flowers/fruitlets 25 days after anthesis and high in developed fruit. Screening the natural variation found in the Israeli germplasm collection revealed that in most cultivars less than one fruit per inflorescence has developed. However, there are unique cultivars with a higher fruit set.     

Factors affecting seed dormancy and germination of Johnsongrass, Sorghum halepense (L.) Pers.

Seeds of Johnsongrass [Sorghum halepense (L.) Pers.] germinated to higher percentages (20–30% higher) when incubated at 28 and 35° C than at 10 or 22° C. After-ripening was accelerated by dry storage of these seeds at 50°C. Seeds pre-chilled at 6°C for 2–4 weeks followed by incubation at 28°C germinated 40–60%. Light effects on germination were related to incubation temperatures; inhibitory at 22°C; no response at 28°C; and stimulatory at 35°C. Effects of gibberellin A₃ (GA₃) also varied depending on incubation temperature, sodium hypochlorite (NaOCl) immersion and light conditions. Immersion of dry seeds in either 700 mM NaOCl, 900 mM H₂O₂ or concentrated H₂SO₄ before incubation in water was effective in breaking dormancy. This result suggests the modes of action of H₂SO₄ in the termination of dormancy may be similar to those of NaOCl and H₂SO₄ as previously suggested by Hsiao & Quick (1984), that is by modification or scarification of the hull or seed coat membranes, and also by the supply of additional oxygen to the seed.     

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.