Descripción: In organello starch biosynthesis was studied using intact chloroplasts isolated from spinach leaves (Spinacia oleracea). Immunoblot analysis using a specific antiserum against the mitochondrial adenylate (ADP/ATP) translocator of Neurospora crassa shows the presence of an adenylate translocator protein in the chloroplast envelope membranes, similar to that existing in mitochondria and amyloplasts from cultured cells of sycamore (Acer pseudoplatanus). The double silicone oil layer-filtering centrifugation technique was employed to study the kinetic properties of adenylate transport in the purified chloroplasts; ATP, ADP, AMP, and most importantly ADP-Glc were shown to be recognized by the adenylate translocator. Similar to the situation with sycamore amyloplasts, only ATP and ADP-Glc uptake was inhibited by carboxyatractyloside, an inhibitor of the mitochondrial adenylate translocator. Evidence is presented to show that the ADP-Glc transported into the chloroplast stroma is utilized for starch synthesis catalyzed by starch synthase (ADP-Glc: 1,4-α-d-glucan 4-α-d-glucosyltransferase). The high activity of sucrose synthase producing ADP-Glc observed in the extrachloroplastic fractions suggests that starch biosynthesis in chloroplasts may be coupled with the direct import of ADP-Glc from the cytosol.

Descripción: Background: Shortly after weaning, a complex multi-step process that leads to massive epithelial apoptosis is triggered by tissue local factors in the mouse mammary gland. Several reports have demonstrated the relevance of mechanical stress to induce adaptive responses in different cell types. Interestingly, these signaling pathways also participate in mammary gland involution. Then, it has been suggested that cell stretching caused by milk accumulation after weaning might be the first stimulus that initiates the complete remodeling of the mammary gland. However, no previous report has demonstrated the impact of mechanical stress on mammary cell physiology. To address this issue, we have designed a new practical device that allowed us to evaluate the effects of radial stretching on mammary epithelial cells in culture. Results: We have designed and built a new device to analyze the biological consequences of applying mechanical stress to cells cultured on flexible silicone membranes. Subsequently, a geometrical model that predicted the percentage of radial strain applied to the elastic substrate was developed. By microscopic image analysis, the adjustment of these calculations to the actual strain exerted on the attached cells was verified. The studies described herein were all performed in the HC11 non-tumorigenic mammary epithelial cell line, which was originated from a pregnant BALB/c mouse. In these cells, as previously observed in other tissue types, mechanical stress induced ERK1/2 phosphorylation and c-Fos mRNA and protein expression. In addition, we found that mammary cell stretching triggered involution associated cellular events as Leukemia Inhibitory Factor (LIF) expression induction, STAT3 activation and AKT phosphorylation inhibition. Conclusion: Here, we show for the first time, that mechanical strain is able to induce weaning-associated events in cultured mammary epithelial cells. These results were obtained using a new practical and affordable device specifically designed for such a purpose. We believe that our results indicate the relevance of mechanical stress among the early post-lactation events that lead to mammary gland involution. © 2009 Quaglino et al., licensee BioMed Central Ltd.

Descripción: We investigate an unsteady plane viscous gravity current of silicone oil on a horizontal glass substrate. Within the lubrication approximation with gravity as the dominant force, this current is described by the nonlinear diffusion equation [Formula Presented]=([Formula Presented][Formula Presented][Formula Presented] (φ is proportional to the liquid thickness h and m=3>0), which is of interest in many other physical processes. The solutions of this equation display a fine example of the competition between diffusive smoothening and nonlinear steepening. This work concerns the so-called waiting-time solutions, whose distinctive character is the presence of an interface or front, separating regions with h≠/0 and h=0, that remains motionless for a finite time interval [Formula Presented] meanwhile a redistribution of h takes place behind the interface. We start the experiments from an initial wedge-shape configuration [h(x)≊[Formula Presented]([Formula Presented]-x)] with a small angle ([Formula Presented]⩽0.12 rad). In this situation, the tip of the wedge, situated at [Formula Presented] from the rear wall (15 cm⩽[Formula Presented]⩽75 cm), waits at least several seconds before moving. During this waiting stage, a region characterized by a strong variation of the free surface slope (corner layer) develops and propagates toward the front while it gradually narrows and [Formula Presented]h/∂[Formula Presented] peaks. The stage ends when the corner layer overtakes the front. At this point, the liquid begins to spread over the uncovered substrate. We measure the slope of the free surface in a range ≊10 cm around [Formula Presented], and, by integration, we determine the fluid thickness h(x) there. We find that the flow tends to a self-similar behavior when the corner layer position tends to [Formula Presented]; however, near the end of the waiting stage, it is perturbed by capillarity. Even if some significant effects are not included in the above equation, the main properties of its solutions are well displayed in the experiments © 1996 The American Physical Society.