Transglutaminase 2 as a biomarker of osteoarthritis: an update

Osteoarthritis is a progressive joint disease characterized by cartilage degradation and bone remodelling. Under physiologic conditions, articular cartilage displays a stable chondrocyte phenotype, whereas in osteoarthritis a chondrocyte hypertrophy develops near the sites of cartilage surface damage and associates to the pathologic expression of type X collagen. Transglutaminases (TGs) include a family of Ca²⁺-dependent enzymes that catalyze the formation of γ-glutamyl cross-links. Their substrates include a variety of intracellular and extracellular macromolecular components. TGs are ubiquitously and abundantly expressed and implicated in a variety of physiopathological processes. TGs activity is modulated by inflammatory cytokines. TG2 (also known as tissue transglutaminase) mediates the hypertrophic differentiation of joint chondrocytes and interleukin-1-induced calcification. Histomorphometrical and biomolecular investigations document increased TG2 expression in human and experimental osteoarthritis. Consequently, the level of TG2 expression may represent an adjuvant additional marker to monitor tissue remodelling occurring in osteoarthritic joint tissue. Experimental induction of osteoarthritis in TG2 knockout mice is followed from reduced cartilage destruction and increased osteophyte formation compared to wild-type mice, suggesting a different influence on joint bone and cartilage remodelling. The capacity of transamidation by TG2 to regulate activation of latent TGF-β seems to have a potential impact on the regulation of inflammatory response in osteoarthritic tissues. Additional studies are needed to define TG2-regulated pathways that are differently modulated in osteoblasts and chondrocytes during osteoarthritis.     

Chemical ecology of Cyclocephala forsteri (Melolonthidae), a threat to macauba oil palm cultivars (Acrocomia aculeata,Arecaceae)

The environmental unsustainability of African oil palm crops is a growing worldwide concern, and the macauba palm in tropical Southern America surges as the most viable substitute for the production of first-generation biodiesel as a renewable fuel. Large-scale cultivation of the macauba palm, however, is bound to suffer a major setback due to the fast emergence of a florivorous pest threat, Cyclocephala forsteri, a scarab beetle species that can severely reduce fruit yield. Like other congenerics, female and male C. forsteri are night-foraging and driven in large numbers to macauba palm inflorescences by specific volatile organic compounds. In the present study, we assessed the kairomonal cues involved in this plant–florivore interaction and their potential application in selective pest management strategies. Headspace samples of the floral fragrance of macauba palms at our study site in Central Brazil were largely dominated by 4-methylanisole (>97% relative amount), which along with the minor constituents 2-isopropyl-3-methoxypyrazine and 2-sec-butyl-3-methoxypyrazine triggered electroantennographic responses from both female and male C. forsteri. Field behavioural assays evidenced that beetles of this species were exclusively attracted to scented traps baited with 4-methylanisole. Increased total scent discharge attained with an ultrasonic piezoelectric diffuser has positively influenced attractiveness efficiency of the trapping setup. 4-Methylanisole is hereby identified as yet another volatile kairomone involved in highly selective attraction of potential pest florivorous cyclocephaline scarabs associated with economically exploitable palms and should be viewed as a key element in short-term integrated management plans for the cultivation of the macauba palm in South America.     

Impact of pesticides in properties of Bradyrhizobium spp. and in the symbiotic performance with soybean

World Journal of Microbiology and Biotechnology - Daptomycin, produced by Streptomyces roseosporus is a novel cyclic lipopeptide antibiotic for treatment of Gram-positive bacteria caused...     

Calcium can moderate changes on membrane structure and lipid composition in cowpea plants under salt stress

The effects of supplemental Ca²⁺ on membrane integrity and lipid composition of cowpea plants submitted to salt stress (75 mM NaCl) were evaluated. The experimental design was factorial (2 × 6 + 1) corresponding to six saline treatments supplemented with CaCl₂ and six saline treatments supplemented with CaSO₄, both at 0.5, 1.25, 2.5, 5.0, 7.5 and 10.0 mM, plus control treatment (plants grown in half-strength Hoagland’s nutrient solution without supplemental calcium addition). Samples of leaves and root tips were analyzed for total lipid, glycolipid and fatty acid contents and membrane damage symptoms. Salt stress greatly reduced total lipid content in leaves and roots and caused great damage to membrane structures. In leaves, the glycolipid content was differently influenced by calcium treatments. Moreover, salinity increased the saturated/unsaturated fatty acid ratio in leaves and an increase in the concentration of calcium intensified this response. In roots, only saturated fatty acids were detected and their content was strongly influenced by salinity and very little by calcium treatments. Supplemental Ca²⁺ was unable to ameliorate the negative effects of salinity on the structural integrity and fluidity of plant membranes in cowpea.     

Differences between epididymal and ejaculated sperm characteristics in donkey

Spermatozoa acquire their motility and fertilizing ability during their passage through the epididymal canal. In the epididymal caput and corpus spermatozoa undergo several biochemical and metabolic changes while the cauda of the epididymis should be considered as the primarily site for storage of the spermatozoa. In the horse spermatozoa from cauda epididymis were collected and frozen, and the fertility of semen assessed. However, no studies have detailed semen characteristics of spermatozoa collected from the cauda epididymis in the jackass. In this study sperm characteristics of spermatozoa in the cauda epididymis of the donkey was reported and a comparison with ejaculated spermatozoal characteristics was performed. Samples from 10 Martina Franca jackasses were collected and analyzed for viability (Propidium iodide/Sybr-14? fluorescent stain), mitochondrial activity (Mitotraker? fluorescent stain), objective motility characteristics (by Computer Assisted Sperm Analyzer - CASA) and morphology. A higher viability and mitochondrial activity in the cauda epididymis samples were reported in this paper. Samples reported in this paper were identified and the percentage of total and progressive spermatozoa was comparable, but trajectories were more rapid (higher VCL) with less progressiveness (higher ALH and lower STR and LIN) in the cauda epididymis. Sperm morphology showed a pronounced variability between jackasses, with comparable values for all morphological subclasses. In this study the loss of the distal cytoplasmic droplets happen close to or after ejaculation because the percentage fell to nearly 0% after ejaculation. As suggested for bulls, the presence of a similar percentage in sperm with proximal cytoplasmic droplet in epididymal and ejaculated semen is likely to indicate a failure in the maturation process.     

Photosynthesis of mistletoes in relation to their hosts at various sites in tropical Brazil

 Chlorophyll a fluorescence parameters showing the instantaneous performance and carbon-isotope ratios reflecting long-term behaviour of leaves were determined for a large number of mistletoe/host-pairs in the cerrado belt of Brazil. Study sites were a very exposed rupestrian field, a semi-exposed savanna and a highly shaded gallery forest. The major question asked was if photosynthetic capacity of mistletoe leaves differed from that of the leaves of their respective hosts. It is shown that except for the very exposed rupestrian field site, photosynthetic capacity appeared to be similar in mistletoes and host leaves. The superior behaviour of host leaves in the rupestrian field was due to particularly expressed sun-plant characteristics of the host. However, mistletoes always had higher average stomatal conductances, lower leaf temperatures at similar or even higher irradiance and higher intercellular CO₂-partial pressures than hosts. Photosynthetic performance of mistletoe leaves was independent of whether a given mistletoe species parasitized aluminium-accumulating or non-accumulating host species in the cerrados with their aluminium-rich soils. Received: 7 April 1997 / Accepted: 20 August 1997     

Human oocytes reversibly arrested in prophase I by phosphodiesterase type 3 inhibitor in vitro

This study addresses the role of cAMP hydrolytic isoenzyme phosphodiesterase type 3 (PDE 3) modulation on human oocyte maturation in vitro. Presence of phosphodiesterase type 3 A (PDE 3A) mRNA was confirmed in human germinal vesicle-stage (GV) oocytes. Making use of a selective PDE 3 inhibitor, Org 9935 (10 microM), oocytes retrieved from immature follicles were arrested in prophase I with a high efficiency for up to 72 h. Cumulus oocyte complexes (COCs) were retrieved in the follicular phase of the cycle before or after exposure to endogenous LH or hCG administration in vivo and randomly distributed into maturation medium with or without the PDE 3 inhibitor. Previous exposure of small follicles to LH activity in vivo had no influence on the arresting capacity of the PDE 3 inhibitor. Reversal from pharmacological arrest leads to a progression through meiosis in a normal time frame with formation of a well-aligned metaphase plate. Ultrastructure analysis of COC derived from follicles between 8 and 12 mm showed that the induced extension of prophase I arrest in vitro resulted in cytoplasm changes but not in apparent nuclear changes during culture.     

Nicotiana benthamiana as a production platform for artemisinin precursors

Background

Production of pharmaceuticals in plants provides an alternative for chemical synthesis, fermentation or natural sources. Nicotiana benthamiana is deployed at commercial scale for production of therapeutic proteins. Here the potential of this plant is explored for rapid production of precursors of artemisinin, a sesquiterpenoid compound that is used for malaria treatment.

Methodology/Principal Findings

Biosynthetic genes leading to artemisinic acid, a precursor of artemisinin, were combined and expressed in N. benthamiana by agro-infiltration. The first committed precursor of artemisinin, amorpha-4,11-diene, was produced upon infiltration of a construct containing amorpha-4,11-diene synthase, accompanied by 3-hydroxy-3-methylglutaryl-CoA reductase and farnesyl diphosphate synthase. Amorpha-4,11-diene was detected both in extracts and in the headspace of the N. benthamiana leaves. When the amorphadiene oxidase CYP71AV1 was co-infiltrated with the amorphadiene-synthesizing construct, the amorpha-4,11-diene levels strongly decreased, suggesting it was oxidized. Surprisingly, no anticipated oxidation products, such as artemisinic acid, were detected upon GC-MS analysis. However, analysis of leaf extracts with a non-targeted metabolomics approach, using LC-QTOF-MS, revealed the presence of another compound, which was identified as artemisinic acid-12-β-diglucoside. This compound accumulated to 39.5 mg.kg⁻¹ fwt. Apparently the product of the heterologous pathway that was introduced, artemisinic acid, is further metabolized efficiently by glycosyl transferases that are endogenous to N. benthamiana.

Conclusion/Significance

This work shows that agroinfiltration of N. bentamiana can be used as a model to study the production of sesquiterpenoid pharmaceutical compounds. The interaction between the ectopically introduced pathway and the endogenous metabolism of the plant is discussed.     

Boophilus microplus Infection by Beauveria amorpha andBeauveria bassiana: SEM Analysis and Regulation of Subtilisin-like Proteases and Chitinases

Beauveria bassiana is a well-known broad-range arthropod pathogen which has been used in biological control of several pest insects and ticks such as Boophilus microplus. Beauveria amorpha has both endophytic and entomopathogenic characteristics, but its capacity for biological control has still not been studied. During the processes of host infection, B. bassiana and B. amorpha produce several hydrolytic extracellular enzymes, including proteases and chitinases, which probably degrade the host cuticle and are suggested to be pathogenicity determinants. To access the role of these enzymes during infection in the tick B. microplus, we analyzed their secretion during fungus growth in single and combined carbon sources, compared to complex substrates such as chitin and B. microplus cuticle. Chitin and tick cuticle-induced chitinase in both fungus and protease was induced only by tick cuticle. SEM analysis of B. amorpha and B. bassiana infecting B. microplus showed apressorium formation during penetration on cattle tick cuticle.