IFI16 induction by glucose restriction in human fibroblasts contributes to autophagy through activation of the ATM/AMPK/p53 pathway

Background

Glucose restriction in cells increases the AMP/ATP ratio (energetic stress), which activates the AMPK/p53 pathway. Depending upon the energetic stress levels, cells undergo either autophagy or cell death. Given that the activated p53 induces the expression of IFI16 protein, we investigated the potential role of the IFI16 protein in glucose restriction-induced responses.

Methodology/Principal Findings

We found that glucose restriction or treatment of human diploid fibroblasts (HDFs) with the activators of the AMPK/p53 pathway induced the expression of IFI16 protein. The induced levels of IFI16 protein were associated with the induction of autophagy and reduced cell survival. Moreover, the increase in the IFI16 protein levels was dependent upon the expression of the functional ATM protein kinase. Importantly, the knockdown of the IFI16 expression in HDFs inhibited the activation of the ATM/AMPK/p53 pathway in response to glucose restriction and also increased the survival of HDFs.

Conclusions/Significance

Our observations demonstrate a role for the IFI16 protein in the energetic stress-induced regulation of autophagy and cell survival. Additionally, our findings also indicate that the loss of IFI16 expression, as found in certain cancers, may provide a survival advantage to cancer cells in microenvironments with low glucose levels.     

Energy exchange and ultrastructure of plant cells under generation of nitric oxide

The effects of nitric oxide (NO) on oxygen consumption, heat generation, and cell ultrastructure were investigated in the seedlings of wheat (Triticum aestivum L.). The experiments were conducted with the excised roots of 5-day-old seedlings grown in the solution of CaCl₂ (2.5 × 10⁻⁴ M). The source of NO was NaNO₂ (5 × 10⁻³ M) where the roots were incubated. Production of NO was determined by means of EPR, respiration — gasometrically, heat generation — using a microcalorimeter. The results showed that NO was formed in the presence of NaNO₂. This was accompanied by a decrease in the respiration rate by about 30%, which lasted for 5–6 h. Apparently, NO inhibited mitochondrial oxidation because stimulation of oxygen consumption induced by 2,4-DNP was completely removed in the presence of NaNO₂. When the cells were affected by succinic acid in the presence of NaNO₂, respiration was strongly inhibited. The effects of succinic acid and NaNO₂ were negated by ascorbic acid. A decrease in the rate of respiration was accompanied by a reduction in heat generation. Moreover, the efflux of potassium ions to the root incubation medium was stimulated, which may point to changes in ionic membrane permeability. The observed changes in energy exchange were accompanied by disturbances in the cell ultrastructure. Nitric oxide induced a clarification of the mitochondrial matrix and a reduction in the number of cristae. It was concluded that NO excess in plant tissues brings about a deceleration of energy exchange, disturbance of the ultrastructural organization, and cell death.     

Automated analysis and classification of infected macrophages using bright-field amplitude contrast data

This article presents a methodology for acquisition and analysis of bright-field amplitude contrast image data in high-throughput screening (HTS) for the measurement of cell density, cell viability, and classification of individual cells into phenotypic classes. We present a robust image analysis pipeline, where the original data are subjected to image standardization, image enhancement, and segmentation by region growing. This work develops new imaging and analysis techniques for cell analysis in HTS and successfully addresses a particular need for direct measurement of cell density and other features without using dyes.     

Changes in the respiratory organs in experimental adenovirus infection

An experiment on 6 green monkeys and on 286 cotton newly born rats was made with the aim of studying the lung during experimental adenovirus infection. All the animals during different terms of infection (from 6 hours and up to 40 days) have been studied. Several morphological changes were discovered in the lungs of monkeys and rats after 6 hours of infection and were retained up to the 10-th day of infection. All the components of air-blood barrier and basal membranes were involved in the process, but after 20 day of the experimental adenovirus infection the entire restoration of cellular structure occurred.     

Peculiarities of functioning of wheat cut-off root cells at inhibition of complexes I and II of mitochondrial respiratory chain

Combined action of rotenone and malonate, inhibitors of complexes I and II of the mitochondrial electron transport chain (ETC), on wheat cut-off root seedlings was studied after 6 h of incubation. Intensity of oxygen consumption and release of potassium ions into incubation medium were determined simultaneously with the study of changes in cell ultrastructure. Malonic acid was added 1 h after the root incubation in the rotenone solution and produced inhibition of respiration, as well as a greater release of K⁺ into the incubation solution as compared with effect of rotenone alone. After 2 h of the combined action of these inhibitors, many mitochondria acquired a toroidal shape, thereby increasing the outer surface. For the ensuing hours, stimulation of oxygen consumption by the roots and a decrease of K⁺ content in the incubation medium were observed. Mitochondria once again acquired a round or oval shape and compensation-reparation processes took place. Contacts of endoplasmic reticulum channels with mitochondria were observed, which seems to be due to the synthesis of the enzyme splitting malonate to acetyl-CoA, which in turn can be included both into the Krebs cycle and into lipogenesis. It is suggested that the toroidal form of mitochondria is associated with the activation of the external NAD(P)H-dehydrogenase of the inner mitochondrial membrane, as under these conditions, at the inhibition of the ETC complexes I and II, the activity of other dehydrogenises is blocked. Thus, the use of the external NAD(P)H allows the activity of the ETC mitochondria to be restored, which facilitates the course of the reparation processes and allows cells to be adapted to this action.     

Spatial and typological arrangement of the population of invertebrates in the grass cover of the Northeastern Altai

The major spatial changes in the communities of invertebrates in the grass cover of the Northeastern Altai and the factors determining these changes are revealed on the basis of multi-year accounts. The strength and general character of the connection between nonhomogeneity of population and environment are estimated.