Detection of conjugated C16 PUFAs in rat tissues as possible partial beta-oxidation products of naturally occurring conjugated linoleic acid and its metabolites

In a previous paper, we showed that naturally occurring conjugated linoleic acid (CLA) from butter fat is metabolized in vivo to higher metabolites such as conjugated diene (CD) 18:3, CD 20:3 and CD 20:4, all the while retaining the conjugated diene structure. In this paper, we describe the detection of two more metabolites with characteristic conjugated diene UV spectra. HPLC retention times, UV and MS spectra identified the CLA metabolites as CD 16:2 and CD 16:3. The accumulation of CD 16:2 was significantly higher than that of CD 16:3 in all tissues examined. Tissue distributions of CD 16:2 and CD 16:3 were similar, with plasma and adipose tissue showing the highest levels, while kidney had the lowest and the liver an intermediate level. CD 16 fatty acids accounted for about 20% of the total CLA metabolites. The kidney, however, was an exception where CD 16 fatty acids accounted for only 11% of total metabolites. Analyses of liver lipid classes showed that CD 16:2 and CD 16:3 were preferentially incorporated into neutral lipids. This preferential incorporation was very similar to CLA as shown previously. We hypothesize that CD 16:2 and CD 16:3 may be derived from partial beta-oxidation of CLA and CD 20:4, respectively, even though we cannot rule out that CD 16:3 may also be derived from CD 18:3 and CD 20:3. Incubation of skin human fibroblasts from X-linked adrenoleukodystrophy (ALD) patients with c9,t11 CLA showed that CD 16:2 formation in ALD cells was about 50% lower than control cells. This result may tempt to hypothesize that, at least in part, CD 16:2 is beta-oxidized in peroxisomes.     

Transplanted mesoangioblasts require macrophage IL-10 for survival in a mouse model of muscle injury

The aim of this study was to verify whether macrophages influence the fate of transplanted mesoangioblasts--vessel-associated myogenic precursors--in a model of sterile toxin-induced skeletal muscle injury. We have observed that in the absence of macrophages, transplanted mesoangioblasts do not yield novel fibers. Macrophages retrieved from skeletal muscles at various times after injury display features that resemble those of immunoregulatory macrophages. Indeed, they secrete IL-10 and express CD206 and CD163 membrane receptors and high amounts of arginase I. We have reconstituted the muscle-associated macrophage population by injecting polarized macrophages before mesoangioblast injection: alternatively activated, immunoregulatory macrophages only support mesoangioblast survival and function. This action depends on the secretion of IL-10 in the tissue. Our results reveal an unanticipated role for tissue macrophages in mesoangioblast function. Consequently, the treatment of muscle disorders with mesoangioblasts should take into consideration coexisting inflammatory pathways, whose activation may prove crucial for its success.     

PCR-based methods to discriminateBacillus thuringiensis strains

Bacillus thuringiensis is a Gram positive bacterium that is used for the production of biopesticides. The toxic action of different strains and serovars ofB. thuringiensis can be extremely selective towards specific pests, or, in contrast, it can affect a wide variety of non-target organisms such as insects, vertebrates or humans. A reliable characterization of the cultivated strains is of primary importance for the biopesticide industry, in order to assess the contamination of the final product with strains with different pesticide actions or that might be dangerous for human health. The aim of this study was to develop useful methods for the typing of differentB. thuringiensis strains using two PCR-based methods, RAPD and Rep-PCR with BOXA1R and ERIC2 primers. The molecular fingerprints obtained using ERIC2-PCR showed a reliable ability to discriminateBacillus thuringiensis strains.     

Study on the conversion of wool keratin by steam explosion

A wool fiber sample was submitted to chemical-free steam explosion in view of potential exploitation of keratin-based industrial and farm wastes. Fiber keratin was converted into a dark-yellow sludge that was submitted to phase separation by filtration, centrifugation, and precipitation of the soluble materials from the supernatant liquid. The resulting products, when compared with the original wool, showed the extent of disruption of the histology structure, reduction of the molecular weight to water-soluble peptides and free amino acids, and change of the structure of the remainder of the protein associated with breaking of disulfide bonds and decomposition of the high-sulfur-content protein fraction.     

Tricarboxylic acid cycle-sustained oxidative phosphorylation in isolated myelin vesicles

The Central Nervous System (CNS) function was shown to be fueled exclusively by oxidative phosphorylation (OXPHOS). This is in line with the sensitivity of brain to hypoxia, but less with the scarcity of the mitochondria in CNS. Consistently with the ectopic expression of F_oF₁-ATP synthase and the electron transfer chain in myelin, we have reported data demonstrating that isolated myelin vesicles (IMV) conduct OXPHOS. It may suggest that myelin sheath could be a site for the whole aerobic degradation of glucose.     

Reversal of the inhibitory action of abscisic acid by benzyladenine in excised watermelon cotyledons

Cotyledons of watermelon ( Citrullus vulgaris Schrad. cv. Fairfax) were excised from the embryo after 24 h of imbibition and cultured for several days on filter paper with water or abscisic acid (ABA) solution. In some experiments the cotyledons were pretreated with benzyladenine (BA) for times ranging from 5 min to 2 h before transfer to ABA.
A treatment with 10⁻⁵ M ABA blocked all developmental parameters examined (growth and increase in appropriate markers for glyoxysome, peroxisome and plastid development). This blocking can be prevented by an initial treatment with 10⁻⁴ M BA for 2 h. This pretreatment with BA overrides the action of ABA: the final developmental responses are not just restored to the level of the water control, but they are almost as high as those obtained by treating the cotyledons with BA only. If BA is administered for three days together with ABA the reversal of inhibition is much less efficient.     

Optically activated and interrogated plasmonic hydrogels for applications in wound healing

We disclose the use of hybrid materials featuring Au/Ag core/shell nanorods in porous chitosan/polyvinyl alcohol scaffolds for applications in tissue engineering and wound healing. The combination of Au and Ag in a single construct provides synergistic opportunities for optical activation of functions as near infrared laser tissue bonding, and remote interrogation to return parameters of prognostic relevance in wound healing monitoring. In particular, the bimetallic component ensures optical tunability, enhanced shelf life and photothermal stability, serves as a reservoir of germicidal silver cations, and changes in near‐infrared and visible color according to the environmental level of oxidative stress. At the same time, the polymeric blend is ideal to bind connective tissue upon photothermal activation, and to support fabrication processes that provide high porosity, such as electrospinning, thus putting all the premises for cellular repopulation and antimicrobial protection.     

Molecular characterization of the human PLC β1 gene

Inositide-specific phospholipase C (PLC) signaling constitutes a central intermediate in a number of cellular functions among which the control of cell growth raises a particular interest. Indeed, we have previously shown that nuclear phospholipase C β1 (PLC β1) is central for the regulation of mitogen-induced cell growth. We have also assigned by fluorescence in situ hybridization (FISH) analysis the PLC β1 to human chromosome 20p12. In this study, we have carried out a detailed analysis of the human gene, showing the existence of alternative splicing, which gives rise, besides the two forms (1a and 1b) already shown in rodents, to a new 600 bp smaller form coding for a 110 kDa protein. We have also identified a new exon at the 5′, showing no homology with the rodent sequence. Here we provide the complete determination of the exon/intron structure of the gene spanning 250 kb of DNA. We found that the exons are quite small, ranging from 49 to 222 bp, while the introns vary between 108 bp and 34,400 bp. The availability of the understanding of the genome organization of this inositide-specific PLC, which represents a key step of the cell cycle related signaling, could actually pave the way for further genetic analysis of p12 region of human chromosome 20 in diseases involving alterations of the control of cell growth such as malignancies.