Store-operated Ca(2+) entry and tyrosine kinase pp60(src) hyperactivity are modulated by hyperglycemia in platelets from patients with non insulin-dependent diabetes mellitus

We have investigated the involvement of store-operated Ca(2+) entry (SOCE) in the abnormal platelet Ca(2+) homeostasis in patients with non insulin-dependent diabetes mellitus (NIDDM). In a medium containing 180 mg/dL glucose, platelets from NIDDM patients showed an increased SOCE compared to controls. We found that tyrosine phosphorylation was elevated in platelets from NIDDM patients. Consistent with this, the activity of the tyrosine kinase pp60(src) is enhanced in platelets from diabetic patients. When the experiments were performed in a medium containing 90 mg/dL both, SOCE and pp60(src) activity, were similar to those found in control platelets. Our results indicate that SOCE is altered in platelets from NIDDM patients probably due to the increased activity of the tyrosine kinase pp60(src). Both, SOCE and pp60(src) activity in platelets from NIDDM patients are more susceptible to the extracellular glucose concentration, which seems to be involved in the dysfunction of these mechanisms.     

Resveratrol mobilizes Ca2+ from intracellular stores and induces c-Jun N-terminal kinase activation in tumoral AR42J cells

Resveratrol (3,4',5-trihydroxy-trans-stilbene), a phytoalexin naturally found in grapes and red wine, is a redox-active compound endowed with significant positive activities. In this study, the effects of resveratrol on intracellular free Ca(2+) concentration ([Ca(2+)](c)) and on cell viability in tumoral AR42J pancreatic cells are examined. The results show that resveratrol (100 μM and 1 mM) induced changes in [Ca(2+)](c), that consisted of single or short lasting spikes followed by a slow reduction toward a value close to the resting level. Lower concentrations of resveratrol (1 and 10 μM) did not show detectable effects on [Ca(2+)](c). Depletion of intracellular Ca(2+) stores by stimulation of cells with 1 nM CCK-8, 20 pM CCK-8 or 1 μM thapsigargin, blocked Ca(2+) responses evoked by resveratrol. Conversely, prior stimulation of cells with resveratrol inhibited Ca(2+) mobilization in response to a secondary application of CCK-8 or thapsigargin. In addition, resveratrol inhibited oscillations in [Ca(2+)](c) evoked by a physiological concentration of CCK-8 (20 pM). On the other hand, incubation of cells in the presence of resveratrol induced a reduction of cell viability. Finally, incubation of AR42J cells in the presence of resveratrol led to activation of c-Jun N-terminal kinase (JNK), a mitogen-activated protein kinase responsive to stress stimuli. Activation of JNK was reduced in the absence of extracellular Ca(2+). In summary, the results show that resveratrol releases Ca(2+) from intracellular stores, most probably from the endoplasmic reticulum, and reduces AR42J cells viability. Reorganization of cell's survival/death processes in the presence of resveratrol may involve Ca(2+)-mediated JNK activation.     

Cinnamtannin B-1, a natural antioxidant that reduces the effects of H<Subscript>2</Subscript>O<Subscript>2</Subscript> on CCK-8-evoked responses in mouse pancreatic acinar cells

This work was designed in order to gain an insight on the mechanisms by which antioxidants prevent pancreatic disorders. We have examined the properties of cinnamtannin B-1, which belongs to the class of polyphenols, against the effect of hydrogen peroxide (H₂O₂) in mouse pancreatic acinar cells. We have studied Ca²⁺ mobilization, oxidative state, amylase secretion, and cell viability of cells treated with cinnamtannin B-1 in the presence of various concentrations of H₂O₂. We found that H₂O₂ (0.1–100 μM) increased CM-H₂DCFDA-derived fluorescence, reflecting an increase in oxidation. Cinnamtannin B-1 (10 μM) reduced H₂O₂-induced oxidation of CM-H₂DCFDA. CCK-8 induced oxidation of CM-H₂DCFDA in a similar way to low micromolar concentrations of H₂O₂, and cinnamtannin B-1 reduced the oxidant effect of CCK-8. In addition, H₂O₂ induced a slow and progressive increase in intracellular free Ca²⁺ concentration ([Ca²⁺]_c). Cinnamtannin B-1 reduced the effect of H₂O₂ on [Ca²⁺]_c, but only at the lower concentrations of the oxidant. H₂O₂ inhibited amylase secretion in response to cholecystokinin, and cinnamtannin B-1 reduced the inhibitory action of H₂O₂ on enzyme secretion. Finally, H₂O₂ reduced cell viability, and the antioxidant protected acinar cells against H₂O₂. In conclusion, the beneficial effects of cinnamtannin B-1 appear to be mediated by reducing the intracellular Ca²⁺ overload and intracellular accumulation of digestive enzymes evoked by ROS, which is a common pathological precursor that mediates pancreatitis. Our results support the beneficial effect of natural antioxidants in the therapy against oxidative stress-derived deleterious effects on cellular physiology.     

Cinnamtannin B-1 as an antioxidant and platelet aggregation inhibitor

Cinnamtannin B-1 is a naturally occurring trimeric A-type proanthocyanidin, present in a limited number of plants, which exhibits a large number of cellular actions mostly derived from its antioxidant properties. Cinnamtannin B-1 modulates several biological processes such as changes in cytosolic free Ca(2+) concentration, endogenous reactive oxygen species generation, protein tyrosine phosphorylation and platelet aggregation. Proanthocyanidins, such as cinnamtannin B-1, have been reported to exert antitumoral activity mediated by a selective proapoptotic action in a number of tumoral cell lines associated with antiapoptotic activity in normal cells. The opposite effects of proanthocyanidins in normal and tumoral cells suggest that these compounds might be the base for therapeutic strategies directed selectively against tumoral cells. In addition, cinnamtannin B-1 shows antithrombotic actions through inhibition, in platelets, of endogenous ROS generation, Ca(2+) mobilization and, subsequently, aggregation. This has been reported to be especially relevant in platelets from diabetic patients, where cinnamtannin B-1 reverses both platelet hypersensitivity and hyperactivity. Considering the large number of cellular effects of cinnamtannin B-1 the development of therapeutic strategies for thrombotic disorders or certain types of cancer deserves further studies. This review summarizes the current knowledge on the actions and relevance of the signalling pathways modulated by cinnamtannin B-1.     

Spalt major controls the development of the notum and of wing hinge primordia of the Drosophila melanogaster wing imaginal disc

The Drosophila wing and the dorsal thorax develop from primordia within the wing imaginal disc. Here we show that spalt major (salm) is expressed within the presumptive dorsal body wall primordium early in wing disc development to specify notum and wing hinge tissue. Upon ectopic salm expression, dorsally located second leg disc cells develop notum and wing hinge tissue instead of sternopleural tissue. Similarly, by salm over-expression within the wing disc, wing blade formation is suppressed and a mirror-image duplication of the notum and wing hinge is formed. In large dorsal clones, which lack salm and its neighboring paralogue spalt related (salr), the cells of the notum primordium do not grow; these dorsal cells are not specified as notum, hence no notum outgrowth develops. These results suggest that the zinc finger factors encoded by the salm/salr complex play important roles in defining cells of the early wing disc as dorsal body wall cells, which develop into a large dorsal body wall territory and form mesonotum and some wing hinge tissue, and in delimiting the wing primordium. We also find that salm activity is down-regulated by its own product and by that of the Pax gene eyegone.     

On the relationship between abiotic stress and co-occurrence patterns: an assessment at the community level using soil lichen communities and multiple stress gradients

The stress‐gradient hypothesis (SGH) predicts a shift from predominant competition to facilitation as abiotic stress increases. Most empirical tests of the SGH have evaluated the interactions between a single or a few pairs of species, have not considered the effects of multiple stress factors, and have not explored these interactions at nested spatial scales. We sampled 63 0.25‐m² plots, each subdivided into 100 5×5 cm and 25 10×10 cm sampling squares, in a semi‐arid Mediterranean environment to evaluate how co‐occurrence patterns among biological soil crusts (BSC)‐forming lichens changed along natural stress gradients driven by water and nutrient (N, P, K) availability. According to the SGH, we tested the hypothesis that the fine‐scale spatial arrangement of BSC‐forming lichens should shift from prevailing interspecific segregation to aggregation as abiotic stress increases. Co‐occurrence patterns ranged from significant species segregation to aggregation at the two spatial scales studied. When using the 5×5 cm grid, more plots showed significant species segregation than aggregation. At this sampling scale, co‐occurrence increased as water and nutrient availability decreased and increased, respectively. Small increases in soil pH promoted species co‐occurrence. Interspecific segregation was promoted as the cover of highly competitive species, such as Diploschistes diacapsis, increased. No significant relationships between co‐occurrence and the surrogates of abiotic stress were observed when data was arranged in a 10×10 cm grid. Our co‐occurrence analyses partially supported predictions from the SGH, albeit the results obtained were dependent on the type of abiotic stress and the spatial scale considered. They show the difficulties of predicting how co‐occurrence patterns change along complex stress gradients, and highlight the need of incorporating the effects of abiotic stress promoted by different resources, such as water and nutrients, into the conceptual framework of the SGH.     

Ruminations on twenty-five years of Patricia Hill Collins's Black Feminist Thought: Knowledge,Consciousness and the Politics of Empowerment

My review focuses on the impact that Black Feminist Thought has had on my personal and professional life. I weave together lessons I have learned from Patricia Hill Collins with reflections on my own lived experience – from my family of origin to college experiences to my work as the founding director of the Collegium of Black Women Philosophers.     

Simulated climate change reduced the capacity of lichen-dominated biocrusts to act as carbon sinks in two semi-arid Mediterranean ecosystems

The importance of biological soil crusts (biocrusts) for the biogeochemistry of drylands is widely recognized. However, there are significant gaps in our knowledge about how climate change will affect these organisms and the processes depending on them. We conducted a manipulative full factorial experiment in two representative dryland ecosystems from central (Aranjuez) and southeastern (Sorbas) Spain to evaluate how precipitation, temperature and biocrust cover affected the assimilation and net C balance of biocrusts. Chlorophyll fluorescence, net photosynthesis and dark respiration were measured in situ bimonthly during a year. We also conducted daily cycle measurements of net photosynthesis in winter and at the end of spring. In Sorbas, warming reduced the fixation of atmospheric C in biocrust-dominated microsites throughout the year. In Aranjuez, there was an interaction between the three factors evaluated; during winter, net photosynthesis was significantly greater in high biocrust cover plots under natural conditions and in the rainfall exclusion treatment. During the daily surveys, rainfall exclusion and warming reduced C fixation in Sorbas and in Aranjuez respectively. The effects of the treatments evaluated varied with the rainfall and non-rainfall water inputs (NRWIs) registered before the measurements. Our results suggest that changes in NRWI regimes as consequence of warming could have a greater impact on the C balance of biocrusts than changes in rainfall amounts. They also indicate that climate change may reduce the photosynthetic ability of lichens, with a consequent reduction of their dominance in biocrust communities at the mid to long term. This could reduce the ability of dryland ecosystems to fix atmospheric C.