The association of insulin resistance with serum osteoprotegerin in obese adolescents

Some data indicate a potential relationship between insulin resistance level and the concentration of osteoprotegerin (OPG) in the body. There have been few studies concerning OPG level and its relationship with insulin resistance and body composition in young people. The aim of this study was to assess serum osteoprotegerin concentration in obese adolescents, and to evaluate its potential association with insulin resistance. Seventy-eight obese adolescents and 20 healthy volunteers aged 12–18 years were recruited in the study. Selected anthropometrical measurements and blood biochemical analyses were performed. Insulin resistance in the participants was evaluated according to the homeostasis model assessment-insulin resistance (HOMA-IR) protocol. Level of OPG was assessed in serum. Obese subjects had significantly higher HOMA-IR indices and OPG levels in serum than the control group. A significant positive correlation between OPG and insulin resistance was found. It was observed that high concentrations of osteoprotegerin are associated with insulin resistance in obese adolescents.     

Casein kinase 1 is a novel negative regulator of E-cadherin-based cell-cell contacts

Cadherins are the most crucial membrane proteins for the formation of tight and compact cell-cell contacts. Cadherin-based cell-cell adhesions are dynamically established and/or disrupted during various physiological and pathological processes. However, the molecular mechanisms that regulate cell-cell contacts are not fully understood. In this paper, we report a novel functional role of casein kinase 1 (CK1) in the regulation of cell-cell contacts. Firstly, we observed that IC261, a specific inhibitor of CK1, stabilizes cadherin-based cell-cell contacts, whereas the overexpression of CK1 disrupts them. CK1 colocalizes with E-cadherin and phosphorylates the cytoplasmic domain of E-cadherin in vitro and in a cell culture system. We show that the major CK1 phosphorylation site of E-cadherin is serine 846, a highly conserved residue between classical cadherins. Constitutively phosphorylated E-cadherin (S846D) is unable to localize at cell-cell contacts and has decreased adhesive activity. Furthermore, phosphorylated E-cadherin (S846D) has weaker interactions with beta-catenin and is internalized more efficiently than wild-type E-cadherin. These data indicate that CK1 is a novel negative regulator of cadherin-based cell-cell contacts.     

The AQEM multimetric system for the southern Italian Apennines: assessing the impact of water quality and habitat degradation on pool macroinvertebrates in Mediterranean rivers

In accordance with the aims of the E.U. funded AQEM Project, an assessment system module based on aquatic macroinvertebrates was developed for small sized rivers in the southern Apennines (south Italy). Eleven stream sites, impacted to a greater or a lesser extent by organic pollution and/or habitat impairment and chosen to cover the whole degradation gradient present in the geographical area were sampled in three seasons. The samples were collected following a proportional, multihabitat procedure, afterwards considering separately the replicates collected in the depositional (pool) and transport (riffle) areas for the analysis. A PCA multivariate analysis was performed to extract the main axes of variation of the biological community, which resulted in the first axis being strongly correlated to ecological quality. The final assessment module is based on a multimetric system, structured by selecting the best metrics in simulating the first axis gradient. The system considers a total of 15 different metrics, mainly providing information concerning tolerance to pollution, taxa richness, habitat features and trophic structure of the community. In accordance with the WFD requirements, some of these metrics are based on abundance classes of taxa. Depositional and transport units, due to the observed dissimilarity in the structure of their benthic communities, were kept separate during the development of the assessment system to retain this potentially useful information and to clear interpretation of the results. Both `riffle' and `pool' invertebrate data showed clear differences in ecological quality between sites. Nevertheless, the final assessment module is based on the macroinvertebrates inhabiting depositional areas of rivers only, because the metrics for these river units showed a better performance than those examined for the transport river units. The application of the assessment module requires 10 replicates to be quantitatively collected, for a total area of 0.5 m². In terms of sampling and identification effort, the assessment module shows a good comparability with the standard Italian method presently in use and might thus be easily applied for river sites classification according to the Water Framework Directive in southern Italy. The site classification obtained with the proposed multimetric index shows a very good correspondence with the post-classification based on multivariate analysis.     

Glycoprotein hypersecretion alters the cell wall in Trichoderma reesei strains expressing the Saccharomyces cerevisiae dolichylphosphate mannose synthase gene

Expression of the Saccharomyces cerevisiae DPM1 gene (coding for dolichylphosphate mannose synthase) in Trichoderma reesei (Hypocrea jecorina) increases the intensity of protein glycosylation and secretion and causes ultrastructural changes in the fungal cell wall. In the present work, we undertook further biochemical and morphological characterization of the DPM1-expressing T. reesei strains. We established that the carbohydrate composition of the fungal cell wall was altered with an increased amount of N-acetylglucosamine, suggesting an increase in chitin content. Calcofluor white staining followed by fluorescence microscopy indicated changes in chitin distribution. Moreover, we also observed a decreased concentration of mannose and alkali-soluble beta-(1,6) glucan. A comparison of protein secretion from protoplasts with that from mycelia showed that the cell wall created a barrier for secretion in the DPM1 transformants. We also discuss the relationships between the observed changes in the cell wall, increased protein glycosylation, and the greater secretory capacity of T. reesei strains expressing the yeast DPM1 gene.     

Quantification of global transcription patterns in prokaryotes using spotted microarrays

We describe an analysis, applicable to any spotted microarray dataset produced using genomic DNA as a reference, that quantifies prokaryotic levels of mRNA on a genome-wide scale. Applying this to Mycobacterium tuberculosis, we validate the technique, show a correlation between level of expression and biological importance, define the complement of invariant genes and analyze absolute levels of expression by functional class to develop ways of understanding an organism's biology without comparison to another growth condition.     

The Characteristics Of Human Bone-Derived Cells (HBDCS) during osteogenesis in vitro

Background

The primary human bone-derived cell culture technique is used as a model to study human osteogenesis. Compared to cell line cultures, primary osteoprogenitor and osteoblast cultures provide more complex information about osteogenesis, bone remodeling and regeneration than cell line cultures.

Methods

In this study, we isolated human bone-derived cells (HBDCs) and promoted their differentiation into osteoblasts. The following parameters were evaluated: cell number and viability, total protein expression, alkaline phosphatase activity, collagenous matrix production and osteogenic genes expression, i.e., gene coding for type I collagen and alkaline phosphatase.

Results

It was proved the results show that HBDCs intensively proliferate during the first 7 days of culture followed by differentiation accompanied by an increase in alkaline phosphatase activity. Moreover, it was observed that during the differentiation of HBDCs, the expression of integrin β1 increased.

Conclusions

The process was also accompanied by changes in cell shape and rearrangement of the actin cytoskeleton and focal contacts containing FAK and the integrin β1 subunit. We suggest that the β1 integrin subunit may be a suitable new target in studies of the differentiation of primary human osteoblasts in culture.

     

Summer diapause in Daphnia as a reaction to the presence of fish

A chemical signal, released by a fish predator under summer-likehigh water temperature and long-day photoperiod, caused theformation of resting eggs in a clone of Daphnia magna. No ephippialfemales were recorded and no ephippia were released in the controltreatment during 45 days of the experiment. When exposed tofish water, the fraction of ephippial females reached a maximumof 3.7%, a value comparable to that registered in summer inthe GroBer Binnensee (Northern Germany), a hypertrophic lakeinhabited by fish, which was the source lake for our experimentalclone. The number of ephippia released within 45 days was onaverage 34 ± 22. Ephippia formation could not resultfrom the between-treatment differences in population density,and related patterns of food depletion, since no substantialdifference between control and fish treatment was observed.Instead, specific information on the presence of a predatorprovided a cue which induced the formation of resting eggs inDaphnia. Under heavy predation and very low survival probabilityof parthenogenetic females, ephippia formation in summer canbe adaptive, i.e. higher fitness can be achieved through survivalin the diapausing state than through the immediate reproductivegain via parthenogenesis.