Hypoxia facilitates the survival of nucleus pulposus cells in serum deprivation by down-regulating excessive autophagy through restricting ROS generation

Nucleus pulposus (NP) cells reside in a hypoxic environment in vivo, while the mechanisms of how NP cells maintain survival under hypoxia are not clear. Autophagy is an important physiological response to hypoxia and implicated in the survival regulation in most types of cells. This study was designed to investigate the role of autophagy in the survival of NP cells under hypoxia. We found that appropriate autophagy activity was beneficial to the survival of NP cells in serum deprivation, while excessive autophagy led to death of the NP cells. Hypoxia facilitated the survival of NP cells in serum deprivation by down-regulating excessive autophagy. Hypoxia down-regulated the autophagy activity of NP cells through restricting the production of reactive oxygen species (ROS) and inactivating the AMPK/mTOR signaling pathway, and possibly through a pathway involving HIF-1α. We believed that understanding the autophagy response of NP cells to hypoxia and its role in cell survival had important clinical significance in the prevention and treatment of degenerative discogenic diseases.     

Endoplasmic reticulum stress in insulin resistance and diabetes

The endoplasmic reticulum is the main intracellular Ca²⁺ store for Ca²⁺ release during cell signaling. There are different strategies to avoid ER Ca²⁺ depletion. Release channels utilize first Ca²⁺-bound to proteins and this minimizes the reduction of the free luminal [Ca²⁺]. However, if release channels stay open after exhaustion of Ca²⁺-bound to proteins, then the reduction of the free luminal ER [Ca²⁺] (via STIM proteins) activates Ca²⁺ entry at the plasma membrane to restore the ER Ca²⁺ load, which will work provided that SERCA pump is active. Nevertheless, there are several noxious conditions that result in decreased activity of the SERCA pump such as oxidative stress, inflammatory cytokines, and saturated fatty acids, among others. These conditions result in a deficient restoration of the ER [Ca²⁺] and lead to the ER stress response that should facilitate recovery of the ER. However, if the stressful condition persists then ER stress ends up triggering cell death and the ensuing degenerative process leads to diverse pathologies; particularly insulin resistance, diabetes and several of the complications associated with diabetes. This scenario suggests that limiting ER stress should decrease the incidence of diabetes and the mobility and mortality associated with this illness.     

Copper exposure reduces production of red carotenoids in a marine copepod

Sub-lethal exposure to copper has been shown to modulate both mitochondrial function and antioxidant gene expression in zooplankton. To date, however, researchers have not identified a quantifiable phenotypic trait that reliably indicates such physiological responses to copper exposure. Red ketocarotenoids are abundant in marine zooplankton serving both physiological and coloration roles, and their production is sensitive to environmental stress. In this study the expression of mitochondrial gene cytochrome c oxidase I (COI) and antioxidant gene glutathione reductase (GR), and the production of red ketocarotenoid, astaxanthin, was measured in response to sub-lethal copper exposure. We found that mRNA of COI and GR was more abundant in copper-exposed copepods than controls, suggesting there was a physiological response to copper exposure. At the same time, copper-exposed copepods produced less astaxanthin than controls. We suggest that ketocarotenoid content of zooplankton has the potential to be a sensitive bioindicator of marine environmental pollution. Understanding how cellular responses to environmental stressors manifest in the phenotypes of marine animals will greatly increase our capacity to monitor marine ecosystem health.     

Herbivory-induced signalling in plants: perception and action

Plants and herbivores have been interacting for millions of years. Over time, plants have evolved mechanisms to defend against herbivore attacks. Herbivore-challenged plants reconfigure their metabolism to produce compounds that are toxic, repellant or anti-digestive for the herbivores. Some compounds are volatile signals that attract the predators of herbivores. All these responses are tightly regulated by a signalling network triggered by the plant's perception machinery. Several compounds that specifically elicit herbivory-induced responses in plants have been isolated from herbivore oral secretions and oviposition fluids. Elicitor perception is rapidly followed by cell membrane depolarization, calcium influx and mitogen-activated protein kinase (MAPK) activation; plants also elevate the concentrations of reactive oxygen and nitrogen species, and modulate phytohormone levels accordingly. In addition to these reactions in the herbivore-attacked regions of a leaf, defence responses are also mounted in unattacked parts of the attacked leaf and as well in unattacked leaves. In this review, we summarize recent progress in understanding how plants recognize herbivory, the involvement of several important signalling pathways that mediate the responses to herbivore attack and the signals that transduce local into systemic responses.     

A combined transcriptional,biochemical and histopathological study unravels the complexity of Alternaria resistance and susceptibility in Brassica coenospecies

Alternaria blight is one of the most devastating diseases of rapeseed-mustard caused by a necrotrophic fungus Alternaria brassicae. Lack of satisfactory resistance resource in Brassica is still a main obstruction for developing resistance against Alternaria. In this study, we have selected Brassica juncea, Sinapis alba and Camelina sativa to understand and unravel the mechanism of disease resistance against Alternaria. Histopathological studies showed early onset of necrosis in B. juncea (1 dpi) and delayed in S. alba (2 dpi) and C. sativa (3 dpi) respectively. Early and enhanced production of hydrogen peroxide (H₂O₂) was observed in C. sativa and S. alba (6 hpi) when compared to B. juncea (12 hpi). An increase in catalase activity was observed in both C. sativa (36 % at 6 hpi) and S. alba (15 % at 12 hpi), whereas it significantly decreased in B. juncea at 6 hpi (23 %), 12 hpi (30 %) and 24 hpi (8 %). Gene expression analysis showed induction of PR-3 and PR-12 genes only in C. sativa and S. alba when compared to B. juncea suggesting their vital role for Alternaria resistance. In contrast, SA marker genes were significantly expressed in B. juncea only which provides evidence of hormonal cross talk in B. juncea during Alternaria infection thereby increasing its susceptibility.     

The Growth,physiological and biochemical response of foxtail millet to atrazine herbicide

Foxtail millet (Pennisetum glaucum L.) is a vital crop that is planted as food and fodder crop around the globe. There is only limited information is present for abiotic stresses on the physiological responses to atrazine. A field experiment was conducted to investigate the effects of different atrazine dosages on the growth, fluorescence and physiological parameters i.e., malonaldehyde (MDA) and reactive oxygen species (ROS) (H₂O₂ and O₂) in the leaves to know the extent of atrazine on oxidative damage of foxtail millet. Our experiment consisted of 0, 2.5, 12.5, 22.5 and 32.5 (mg/kg) of labeled atrazine doses on 2 foxtaill millet varieties. High doses of atrazine significantly enhanced ROS and MDA synthesis in the plant leaves. Enzymes activities like ascorbate peroxidase (APX) and peroxidase (POD) activities enhanced, while catalase (CAD) and superoxide dismutase (SOD) activities reduced with increasing atrazine concentrations. Finally atrazine doses at 32.5 mg/kg reduced chlorophyll contents, while chlorophyll (a/b) ratio also enhanced. Biomass, plant height, chlorophyll fluorescence parameters, minimal and maximal fluorescence (Fo, Fm), maximum and actual quantum yield, photochemical quenching coefficient, and electron transport rate are decreased with increasing atrazine doses.     

Analysis of native and oxidized low-density lipoprotein oxysterols using gas chromatography—mass spectrometry with selective ion monitoring

SUMMARY

Cholesterol oxidation products have been demonstrated to possess a wide variety of biological properties and have been implicated in playing an important role in the development of atherosclerosis. We have developed an analytical method using capillary gas chromatography-mass spectrometry (GC-MS) for the analysis of cholesterol oxidation products in low-density lipoprotein (LDL). The method uses programmed multiple selected ion monitoring (SIM), providing enhanced sensitivity and accuracy of peak detection over full-scan mass spectra. The major oxidation products of cholesterol in oxidized LDL were identified as 7β-hydroxy-cholesterol and 7-keto-cholesterol. Minor products included 4β-hydroxy-cholesterol, 6β-hydroxy-cholesterol and cholesterol-5α,6α-epoxide. Native LDL contains 7-lathosterol, which is a biosynthetic precursor of cholesterol, as well as low levels of 7β-hydroxy-cholesterol and 7-keto-cholesterol. 7-Lathosterol was not detected in oxidized LDL. A time course oxidation of native LDL with 8 μM CuCl₂ demonstrated a rapid increase in 7β-hydroxy-cholesterol and 7-keto-cholesterol over the first 4 h. Cholesterol—5α,6α-epoxide, and β4-hydroxy- and 6β-hydroxy-cholesterol levels increased gradually, while 7-lathosterol decreased over the same period. This method was used to measure the levels of 7-lathosterol and cholesterol oxides in the LDL of 20 healthy subjects in order to establish the mean concentration and a reference range. This method can be used for the characterization and quantitation of oxysterols in native and oxidized LDL and may afford an additional index of oxidative modification of plasma lipoproteins.     

Glutathione Restricts Serine Metabolism to Preserve Regulatory T Cell Function

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磷酸肌醇激酶FAB1调控拟南芥根毛伸长

FAB1/PIKfyve是介导PI(3,5)P₂ (磷脂酰肌醇3,5-二磷酸)生物合成的磷酸肌醇激酶。在动物和酵母(Saccharomyces cerevisiae)中, PI(3,5)P₂参与调控胞内膜运输, 但在植物中的研究较少。该文通过分析拟南芥(Arabidopsis thaliana) FAB1的T-DNA插入突变体的表型解析PI(3,5)P₂的生物学功能。拟南芥FAB1基因家族包含FAB1A、FAB1B、FAB1C和FAB1D四个基因。研究发现, fab1a/b呈现雄配子体致死的表型。利用遗传杂交获得fab1b/c/d三突变体, 发现FAB1B、FAB1C和FAB1D功能缺失导致根毛相比野生型变短, 经FAB1特异性抑制剂YM201636处理后的野生型中也观察到相似的短根毛表型。此外, fab1b/c/d三突变体中DR5转录水平降低。同时, 外源施加生长素类似物2,4-D和NAA能部分恢复fab1b/c/d植株短根毛的表型, 但fab1b/c/d突变体对生长素转运抑制剂(1-NOA和TIBA)的敏感性与野生型相似。此外, FAB1B/C/D功能缺失使根毛中ROS的含量减少且影响肌动蛋白的表达。上述结果表明, FAB1B/C/D通过调控生长素分布、ROS含量和肌动蛋白的表达影响拟南芥根毛伸长。