Proteomic and redox‐proteomic evaluation of homogentisic acid and ascorbic acid effects on human articular chondrocytes

Alkaptonuria (AKU) is a rare genetic disease associated with the accumulation of homogentisic acid (HGA) and its oxidized/polymerized products in connective tissues up to the deposition of melanin‐like pigments (ochronosis). Since little is known on the effects of HGA and its metabolites on articular cells, we carried out a proteomic and redox‐proteomic analysis to investigate how HGA and ascorbic acid (ASC) affect the human chondrocytic protein repertoire. We settled up an in vitro model using a human chondrocytic cell line to evaluate the effects of 0.33 mM HGA, alone or combined with ASC. We found that HGA and ASC significantly affect the levels of proteins with specific functions in protein folding, cell organization and, notably, stress response and cell defense. Increased protein carbonyls levels were found either in HGA or ASC treated cells, and evidences produced in this paper support the hypothesis that HGA‐induced stress might be mediated by protein oxidation. Our finding can lay the basis towards the settling up of more sophisticated models to study AKU and ochronosis. J. Cell. Biochem. 111: 922–932, 2010. © 2010 Wiley‐Liss, Inc.     

Elemental variability in the coralline alga Lithophyllum yemenense as an archive of past climate in the Gulf of Aden (NW Indian Ocean)

This study presents the first algal thallus (skeleton) archive of Asian monsoon strength and Red Sea influence in the Gulf of Aden. Mg/Ca, Li/Ca, and Ba/Ca were measured in Lithophyllum yemenense from Balhaf (Gulf of Aden) using laser ablation inductively coupled plasma mass spectrometry, and Mg/Ca ratio oscillation was used to reconstruct the chronology (34 y). Oscillations of element rates corresponding to the algal growth between 1974 and 2008 were compared with recorded climate and oceanographic variability. During this period, sea surface temperatures (SST) in Balhaf recorded a warming trend of 0.55°C, corresponding to an increase in Mg and Li content in the algal thallus of 2.1 mol‐% and 1.87 μmol‐%, respectively. Lithophyllum yemenense recorded decadal SST variability by Li/Ca, and the influence of the Pacific El‐Niño Southern Oscillation on the NW Indian Ocean climate system by Ba/Ca. Additionally, algal Mg/Ca, Li/Ca, and Ba/Ca showed strong and significant correlations with All Indian Rainfall in the decadal range indicating that these proxies can be useful for tracking variability in the Indian monsoon system, possibly due to changes of the surface wind system, with deep water upwelling in summer, and a distinct seasonality.     

Effect of the [CCTG]n repeat expansion on ZNF9 expression in myotonic dystrophy type II (DM2)

Myotonic dystrophy is caused by two different mutations: a (CTG)n expansion in 3' UTR region of the DMPK gene (DM1) and a (CCTG)n expansion in intron 1 of the ZNF9 gene (DM2). The most accredited mechanism for DM pathogenesis is an RNA gain-of-function. Other findings suggest a contributory role of DMPK-insufficiency in DM1. To address the issue of ZNF9 role in DM2, we have analyzed the effects of (CCTG)n expansion on ZNF9 expression in lymphoblastoid cell lines (n=4) from DM2 patients. We did not observe any significant alteration in ZNF9 mRNA and protein levels, as shown by QRT-PCR and Western blot analyses. Additional RT-PCR experiments demonstrated that ZNF9 pre-mRNA splicing pattern, which includes two isoforms, is unmodified in DM2 cells. Our results indicate that the (CCTG)n expansion in the ZNF9 intron does not appear to have a direct consequence on the expression of the gene itself.     

A proteomic study on human osteoblastic cells proliferation and differentiation

Changes in expression profiles for 17 proteins were ascertained in human mature osteoblasts compared to pre-osteoblasts (differentiation markers). A differential approach was used to highlight proteomic changes between human osteosarcoma cells and mature osteoblasts, showing a relative over-expression of 8 proteins (proliferation and tumor indicators), as well as under-expression of proteins also found down-regulated in pre-osteoblasts (specific markers of osteoblast differentiation). Our findings confirmed the differences between cell lines and primary human cell cultures and suggested caution on the use of osteosarcoma to study anti-osteoporotic drugs in humans.     

Cross-linking of the mannose receptor on monocyte-derived dendritic cells activates an anti-inflammatory immunosuppressive program

Immature monocyte-derived dendritic cells (DC) strongly express the endocytic mannose receptor (MR). Addition of a specific anti-MR mAb (clone PAM-1) for 24 h to cultures of immature DC induced phenotypical and functional maturation of the cells, assessed as up-regulation of costimulatory molecules and CD83, and chemotactic response to CCL19. A different isotype-matched anti-MR mAb (clone 19.2) had no significant effect. Engagement of MR with mAb PAM-1 induced the production of the anti-inflammatory cytokines IL-10, IL-1R antagonist, and of the nonsignaling IL-1R type II. In contrast IL-1beta, TNF, and IL-12 were not produced. PAM-1-treated DC were unable to polarize Th1 effector cells and did not secrete the chemokines CXCL10 and CCL19; in turn, they produced large amounts of CCL22 and CCL17, thus favoring the amplification of Th2 circuits. T cells cocultured with PAM-1-matured DC initially proliferated but later became anergic and behaved as suppressor/regulatory cells. Natural ligands binding to MR had differential effects. MUC III (a partially purified mucin), biglycan (a purified complex proteoglycan), and mannosylated lipoarabinomannan from Mycobacterium tuberculosis affected cytokine production with high IL-10, IL-1R antagonist, IL-1R type II, and inhibition of IL-12. In contrast, mannan, dextran, and thyroglobulin had no significant effect. In conclusion, the appropriate engagement of the MR by mAb PAM-1 and selected natural ligands elicit a secretory program in mono-derived DC characterized by a distinct profile of cytokines/chemokines with the ability to dampen inflammation and to inhibit the generation of Th1-polarized immune responses.     

Effect of culture age, protectants, and initial cell concentration on viability of freeze-dried cells of Metschnikowia pulcherrima

The effect of freeze-drying using different lyoprotectants at different concentrations on the viability and biocontrol efficacy of Metschnikowia pulcherrima was evaluated. The effects of initial yeast cell concentration and culture age on viability were also considered. Yeast cells grown for 36 h were more resistant to freeze-drying than were 48 h cells. An initial concentration of 10? cells·mL?1 favoured the highest survival after freeze-drying. When maltose (25%, m/v) was used as protectant, a high cell viability was obtained (64.2%). Cells maintained a high viability after 6 months of storage at 4 °C. The biocontrol efficacy of freeze-dried cells was similar to the activity of fresh cells on 'Gala' apples and was slightly lower on 'Golden Delicious' apples. After optimizing freeze-drying conditions, the viability of M. pulcherrima cells was similar to that obtained in other studies. The results constitute a first step towards the commercial development of M. pulcherrima as a biocontrol agent.     

Prognostic potential of precise molecular diagnosis of Autosomal Recessive Osteopetrosis with respect to the outcome of bone marrow transplantation

Hematopoietic stem cell transplantation (HSCT) is often the only practical approach to fatal genetic defects. One of the first pathologies which HSCT was applied to was Autosomal Recessive Osteopetrosis (ARO), a rare genetic bone disease in which a deficit in bone resorption by osteoclasts leads to increased bone density and secondary defects. The disease is often lethal early in life unless treated with HSCT. In utero transplantation (IUT) of the oc/oc mouse, reproducing the clinical features of a subset of ARO, has demonstrated that the quality of life and the survival of transplanted animals are greatly improved, suggesting that a similar protocol could be applied to humans. However, recently the dissection of the molecular bases of the disease has shown that ARO is genetically heterogeneous and has revealed the presence of subsets of patients which do not benefit from HSCT. This observation highlights the importance of molecular diagnosing ARO to identify and establish the proper therapies for a better prognosis. In particular, on the basis of experimental results in murine models, efforts should be undertaken to develop approaches such as IUT and new pharmacological strategies.     

Haemagglutination and surface structures in strains of Clostridium spiroforme

The low heat resistance (D100 = 0.554 min, z = 13.4 degrees C) of dormant lysozyme-sensitized spores of Bacillus sphaericus 9602 was correlated with a low protoplast wet density (1.305 g/ml) equivalent to a high protoplast water content (61.0%, wet weight basis). These values for these unusual spores were consistent with those correlated previously in 28 spore types of seven other species.     

Salinity-induced limits to mangrove canopy height

Aim

Mangrove canopy height is a key metric to assess tidal forests' resilience in the face of climate change. In terrestrial forests, tree height is primarily determined by water availability, plant hydraulic design, and disturbance regime. However, the role of water stress remains elusive in tidal environments, where saturated soils are prevalent, and salinity can substantially affect the soil water potential.

Location

Global.

Time Period

The canopy height dataset provides a global snapshot of the maximum mangrove height geographical distribution for the year 2000. Climate and environmental variables extend over the period 1970–2018.

Major Taxa Studied

Mangroves.

Methods

We use global observations of maximum canopy height, species richness, air temperature, and seawater salinity—a proxy of soil water salt concentration—to explore the causal link between salinity and mangrove stature.

Results

Our findings suggest that salt stress limits mangrove height. High salinity favours more salt-tolerant species, narrowing the spectrum of viable traits. Highly salt-tolerant mangroves have evolved to cope with high salt concentrations in the soil, but this adaptation comes at a cost. They typically have lower rates of photosynthesis and growth, resulting in reduced productivity and smaller stature compared to more salt-sensitive mangrove species. This suggests a causal link between salinity, biodiversity, and tree height, where high salinity selects for more salt-tolerant species that tend to be less productive and shorter.

Conclusions

We hypothesize that the salinity-induced limit to mangrove canopy height is the direct result of a reduction of primary productivity, an increment in the risk of xylem cavitation, and an indirect consequence of the decrease in biodiversity. As sea-level rise enhances coastal salinisation, failure to account for these effects can lead to incorrect estimates of future carbon stocks in Tropical coastal ecosystems and endanger preservation efforts.