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.     

Methanolic extract of Verbascum macrurum as a source of natural preservatives against oxidative rancidity

The antioxidant properties of various fractions of a methanolic extract obtained from the aerial parts of Verbascum macrurum have been determined by monitoring their capacity to scavenge the stable free-radical DPPH. They were also evaluated as natural preservatives against oxidative rancidity using the accelerated Rancimat method. Their activities expressed as protection factor (PF(r)) indicated that the fractions rich with phenylpropanoid glycosides were more potent compared to alpha-tocopherol and of the same magnitude as BHT, which were used as reference standards. Ten natural compounds were identified as components of this methanolic extract and were isolated by medium-pressure liquid chromatography (MPLC). Assessment of their antioxidant activities established that acteoside, a polyhydroxylated phenylpropanoid glycoside derivative, is the most potent free radical scavenger and showed the highest protection factor (PF(r)) against sunflower-oil-induced oxidative rancidity. Its activity is comparable to the synthetic antioxidant BHT and clearly superior to natural alpha-tocopherol. This compound therefore represents a very interesting candidate for use in food preservation as natural protecting agent against oxidative rancidity.     

Decoding attraction: Improving vine weevil monitoring by exploiting key sensory cues

BACKGROUND

Monitoring is an integral component of integrated pest management (IPM) programmes used to inform crop management decisions. Vine weevil, Otiorhynchus sulcatus F. (Coleoptera: Curculionidae), continues to cause economically significant losses in horticultural crops due to an inability to reliably detect the presence of this species before crop damage occurs. To improve vine weevil monitoring we investigated the behavioural responses of adult vine weevils to visual (monitoring tool shade/colour, height and diameter as well as the effect of monitoring tool and plant density) and olfactory (host plant and conspecifics) cues under glasshouse conditions.

RESULTS

Monitoring tool shade, height and diameter all influenced monitoring tool efficacy, with individuals exhibiting a preference for black, tall and wide monitoring tools. The total number of individuals recorded in monitoring tools increased with monitoring tool density. By contrast, plant density did not influence the number of individuals recorded in monitoring tools. Yew-baited monitoring tools retained a larger number of individuals compared to unbaited ones. Similarly, more vine weevils were recorded in monitoring tools baited with yew and conspecifics than in unbaited monitoring tools or those baited with only yew. Baiting monitoring tools with conspecifics alone did not enhance the number of vine weevils recorded in monitoring tools.

CONCLUSIONS

Our study confirms that visual and olfactory cues influence vine weevil behaviour. This provides information on key factors that influence vine weevil monitoring tool efficacy and can be used to inform the development of a new monitoring tool for this pest. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Do pine plantations provide mycorrhizal inocula for seedlings establishment in grasslands from Patagonia,Argentina?

We investigated if Pinus ponderosa plantations in Patagonia are able to produce viable mycorrhizal inocula towards adjacent grasslands, which only harbor endomycorrhizal vegetation. We hypothesized that these inocula have the potential to contribute to the establishment of naturally disseminated seedlings. Also, we determined the main fungal taxa involved in this process. Seven plantations in the onset of their reproductive phase and located in the Patagonian native forest/steppe ecotone (Argentina) were selected. Soil samplings were obtained at nine points along a 450 m long, W-E transect established in each plantation. Soil bioassays were performed in a greenhouse, with P. ponderosa seedlings acting as hosts for mycorrhizal inocula present in soil samples, during 12 months. Mycorrhization percentage, morphotype richness and morphotype composition was determined through morphological evaluation. Viable ecto- and ectendomycorrhizal inocula were found disseminated outside plantations. The amount of mycorrhizal inoculum followed a decreasing function with distance to plantation edges. Mycorrhizal fungal genus Rhizopogon and “E-strain” mycorrhizal types appeared as pioneering taxa regarding seedlings colonization, being the most persistent and frequent symbionts found. Plantations, thus, facilitate the surrounding terrain for newcoming seedlings through the dispersion of mycorrhizal fungal inocula.     

Food additive lactic acid production by immobilized cells of Lactobacillus brevis on delignified cellulosic material

Improvements in yield and productivity in lactic acid fermentation by Lactobaccilus brevis cells immobilized on delignified cellulosic (DC) material are reported. The system proved to be more efficient in comparison with the work reported by other workers. Yields of 80 and 100% conversion using glucose were obtained at 30 degrees C in 1 day of fermentation time. Lactic acid fermentation using whey as substrate was obtained at 30 degrees C in 1-1.5 days, resulting in 70% yield, whereas the remaining lactose in whey was converted to alcohol byproduct, leading to a 90% lactose exploitation and 100% conversion. Cell immobilization of L. brevis on DC material was proved by its reuses in repeated batch fermentations and through electron microscopy. A series of 10 repeated batch fermentations without any loss in cell activity showed a tendency for high operational stability. The presence of DC material resulted in a drastic drop of the fermentation time from 48 to 13 h.     

Oestradiol Induced Inhibition of Neuroendocrine Marker Expression in Leydig Cells of Adult Rats

The objectives of this work were to determine the changes in the expression of neuroendocrine markers in Leydig cell by oestradiol treatment, and to determine whether testosterone is able to recover partially the effects of hormonal suppression induced by oestradiol. Adult male rats were injected daily with either 50 microg of oestradiol or oestradiol plus testosterone propionate (25 mg every 3 days) for 15 days. The animals were sacrificed and testicles were dissected and processed by routine histological protocols. FSH and LH serum levels were determined by radioimmunoassay. The visualization of antigens was achieved by the streptavidin-peroxidase immunohistochemical method. Antibodies against chromogranin A (CrA), S-100 protein (S-100), P substance (PS), synaptofisin (SYN), neurofilament protein (NF), gliofibrillary acidic protein (GFAP) and neuron specific enolase (NSE) were used. The mean LH and FSH serum concentrations were consistently suppressed with hormonal treatments. Intermediate filaments (NF and GFAP) showed no difference in their expression. The expression of S-100, NSE and SYN was significantly lower in both hormone-treated groups. In oestradiol-treated rats, the immunoreactivity of CrA and SP decreased significantly but was restored after testosterone supplementation. Although the nature and functions of many of these substances in Leydig cells remain unknown, these results are consistent with the hypothesis that the expression of some neuroendocrine markers is hormonally controlled.     

Management of feral domestic cats in the urban environment of Rome (Italy)

In Italy, which is rabies-free, the national Law No. 281 [Legge Nazionale 14 agosto 1991. No. 281: Legge Quadro in materia di animali di affezione e prevenzione del randagismo. Gazz. Uff. Rep. Ital. no 203 del 30 agosto 1991: p. 3] on the management of pets and on the control of feral cats has introduced the no-kill policy for this species. Thus, “trap-neuter-release” (TNR) programs have been carried out for >10 years. In this paper we present data on registered colonies and censused cats in Rome from 1991 to 2000; the results of the neutering campaign from 1991 to 2000; and a survey, on 103 cat colonies, on the effects of demographic control of urban feral-cat colonies in the city of Rome, carried out by the local Veterinary Public Services (VPS) in collaboration with the associations of cat care-takers. In 10 years almost 8000 were neutered and reintroduced in their original colony. The spay/neuter campaigns brought about a general decrease in cat number but the percentage of cat immigration (due to abandonment and spontaneous arrival) is around 21%. This suggests that all these efforts without an effective education of people to control the reproduction of house cats (as a prevention for abandonment) are a waste of money, time and energy.     

Genotoxic activity of dichlorvos,trichlorfon and dichloroacetaldehyde

Two structurally related organophosphorous insecticides, dichlorvos and trichlorfon, and their main metabolite dichloroacetaldehyde, were assayed for their ability to induce DNA repair, detected as unscheduled DNA synthesis in human epithelial-like cell (EUE) cultures in vitro. A dose-response relationship was found for treatment with the two pesticides, but negative results were obtained with dichloroacetaldehyde. Tests for induction of gene mutation, as ouabain resistance in Chinese hamster cells (V79), failed to show any mutagenic activity by these compounds.     

Characterization of the cytokine pattern of porcine bone marrow-derived cells treated with 1alpha,25(OH)D

The biologically active form of vitamine D(3) [1alpha,25(OH)(2)D(3)] has recently been described not only to influence bone metabolism but also to exert immunomodulating activities, which may have an impact on bone formation/resorption as well. In this study, we analysed the effects of 1alpha,25(OH)(2)D(3) on the cytokine pattern of porcine bone marrow-derived cells from piglets aged 1-3 weeks. After culture for 1 week, the number of osteoclasts was determined, with tartrate-resistant acid phosphatase (TRAP)-positive, multinucleated cells being considered osteoclasts. Cultured bone marrow cell-derived mRNA was subjected to semiquantitative RT-PCR specific for a panel of porcine cytokines (IL-1alpha, IL-6, IL-8, IL-10, and TNF-alpha). In addition, an immunofluorescence analysis using anti-porcine mAbs specific for IL-1beta, IL-2, IL-4, IL-6, IL-12, TNF-alpha, and IFN-gamma was performed. In order to prove the existence of a porcine homologue of the receptor activator of NF-kappaB ligand (RANKL) bone marrow cell- as well as porcine white blood cell-derived mRNA was investigated by RT-PCR using primer pairs specific for murine RANKL. Cell culture supernatant was analysed for soluble RANKL by means of an ELISA designed for quantification of human RANKL. By means of RT-PCR, expression of IL-1alpha, IL-6, IL-8, IL-10 and TNF-alpha mRNA could be found in cells cultured with and without 1alpha,25(OH)(2)D(3). Immunofluorescence analysis revealed that IL-1, IL-6, and TNF-alpha were produced by both stromal cells and osteoclasts. Besides its known osteoclastogenic effects, 1alpha,25(OH)(2)D(3) tended to downregulate the respective cytokines, but significantly upregulated RANKL expression. The homology between the porcine RANKL-specific sequence and the corresponding human RANKL sequence was 79%. The data found support the idea that porcine bone marrow cell cultures may provide a suitable alternative to murine systems in human osteological research.