Tissue processing of nitrite in hypoxia: an intricate interplay of nitric oxide-generating and -scavenging systems

Although nitrite (NO(2)(-)) and nitrate (NO(3)(-)) have been considered traditionally inert byproducts of nitric oxide (NO) metabolism, recent studies indicate that NO(2)(-) represents an important source of NO for processes ranging from angiogenesis through hypoxic vasodilation to ischemic organ protection. Despite intense investigation, the mechanisms through which NO(2)(-) exerts its physiological/pharmacological effects remain incompletely understood. We sought to systematically investigate the fate of NO(2)(-) in hypoxia from cellular uptake in vitro to tissue utilization in vivo using the Wistar rat as a mammalian model. We find that most tissues (except erythrocytes) produce free NO at rates that are maximal under hypoxia and that correlate robustly with each tissue's capacity for mitochondrial oxygen consumption. By comparing the kinetics of NO release before and after ferricyanide addition in tissue homogenates to mathematical models of NO(2)(-) reduction/NO scavenging, we show that the amount of nitrosylated products formed greatly exceeds what can be accounted for by NO trapping. This difference suggests that such products are formed directly from NO(2)(-), without passing through the intermediacy of free NO. Inhibitor and subcellular fractionation studies indicate that NO(2)(-) reductase activity involves multiple redundant enzymatic systems (i.e. heme, iron-sulfur cluster, and molybdenum-based reductases) distributed throughout different cellular compartments and acting in concert to elicit NO signaling. These observations hint at conserved roles for the NO(2)(-)-NO pool in cellular processes such as oxygen-sensing and oxygen-dependent modulation of intermediary metabolism.     

The human phosphatase interactome: An intricate family portrait

The concerted activities of kinases and phosphatases modulate the phosphorylation levels of proteins, lipids and carbohydrates in eukaryotic cells. Despite considerable effort, we are still missing a holistic picture representing, at a proteome level, the functional relationships between kinases, phosphatases and their substrates. Here we focus on phosphatases and we review and integrate the available information that helps to place the members of the protein phosphatase superfamilies into the human protein interaction network. In addition we show how protein interaction domains and motifs, either covalently linked to the phosphatase domain or in regulatory/adaptor subunits, play a prominent role in substrate selection.     

模拟海拔3600m低氧环境对人的认知灵活性的影响

目的：探讨模拟高原低氧环境对人认知灵活性的影响。方法：低氧舱模拟海拔3600m低氧环境,采用任务转换范式观察低氧模拟各阶段的认知灵活性,同时监测焦虑状态及基本生理指标的变化。23名无高原生活经验、平均年龄25．1岁的男性受试者参加了实验。结果：与低氧暴露1个月后的基础对照相比,低氧阶段的反应时转换损失显著增加;低氧阶段焦虑水平显著高于适应阶段;在适应阶段,焦虑水平与反应时转换损失显著负相关;而在降舱阶段,焦虑水平与反应时转换损失显著正相关。结论：中度低氧暴露影响人的认知灵活性和焦虑状态;低氧暴露前,焦虑可促进个体的认知灵活性;低氧暴露后,焦虑会阻碍个体的认知灵活性。     

Endotoxin-induced liver hypoxia: defective oxygen delivery versus oxygen consumption.

In vivo EPR was used to investigate liver oxygenation in a hemodynamic model of septic shock in mice. Oxygen-sensitive material was introduced either (i) as a slurry of fine particles which localized at the liver sinusoids (pO2 = 44.39 +/- 5.13 mmHg) or (ii) as larger particles implanted directly into liver tissue to measure average pO2 across the lobule (pO2 = 4.56 +/- 1.28 mmHg). Endotoxin caused decreases in pO2 at both sites early (5-15 min) and at late time points (6 h after endotoxin; sinusoid = 11.22 +/- 2.48 mmHg; lobule = 1.16 +/- 0.42 mmHg). The overall pO2 changes observed were similar (74.56% versus 74.72%, respectively). Blood pressures decreased transiently between 5 and 15 min (12.88 +/- 8% decrease) and severely at 6 h (59 +/- 9% decrease) following endotoxin, despite volume replacement with saline. Liver and circulatory nitric oxide was elevated at these times. Liver oxygen extraction decreased from 44% in controls to only 15% following endotoxin, despite severe liver hypoxia. Arterial oxygen saturation, blood flow (hepatic artery), and cardiac output were unaffected. Pretreatment with l-NMMA failed to improve endotoxin-induced oxygen defects at either site, whereas interleukin-13 preserved oxygenation. These site-specific measurements of pO2 provide in vivo evidence that the principal cause of liver hypoxia during hypodynamic sepsis is reduced oxygen supply to the sinusoid and can be alleviated by maintaining sinusoidal perfusion.     

Intermediary metabolism: An intricate network at the crossroads of cell fate and function

Intermediary metabolism is traditionally viewed as the large, highly integrated network of reactions that provides cells with metabolic energy, reducing power and biosynthetic intermediates. The elucidation of its major pathways and molecular mechanisms of energy transduction occupied some of the brightest scientific minds for almost two centuries. When these goals were achieved, a sense that intermediary metabolism was mostly a solved problem pervaded the broader biochemical community, and the field lost its vitality. However, intermediary metabolism has recently been re-energized by several paradigm-shifting discoveries that challenged its perception as a self-contained system and re-positioned it at the crossroads of all aspects of cell function, from cell growth, proliferation and death to epigenetics and immunity. Emphasis is now increasingly placed on the involvement of metabolic dysfunction in human disease. In this review, we will navigate from the dawn of intermediary metabolism research to present day work on this ever-expanding field.     

Effect of high altitude hypoxia on the human EEG

The paper presents the results of the comparative study of the EEG at alpine altitudes (Tuya -- Ashu pass, 3200 m) and at low altitudes (City of Frunze, 760 m above the sea level). The dynamics of EEG changes at different stages of adaptation to hypoxia is also traced. The obtained data show that the alpine hypoxia produces a considerable intensification of the excitation processes in the cerebral cortex. Different sensitivity to the oxigen shortage has been found in the frontal-temporal parts of the right and the left hemispheres.     

模拟高原低氧对人的认知能力影响的研究

目的 :研究轻、中度急性缺氧暴露对人的认知能力的影响。方法 :利用低压舱模拟 30 0m(对照高度 )、2 80 0m、36 0 0m、44 0 0m低氧环境 ,考察了 18名健康男性青年受试者在不同高度暴露 1h的感知觉、记忆、注意、思维和心理运动功能的变化情况。结果 :2 80 0m暴露 1h连续识别记忆测验的综合绩效已显著低于对照水平 (P <0 .0 1) ,而36 0 0m暴露 1h后感知觉、记忆、注意、思维和心理运动等各项测验的反应时均延长 ,正确率下降 ,综合绩效显著降低 (P <0 .0 5或P <0 .0 1) ,并随着高度的增加而加重。 44 0 0m暴露 1h各项测验综合绩效进一步降低 (P <0 .0 1)。结论 :各项认知能力受低氧影响符合“高度依赖原则” ,但不同认知能力要素受低氧影响的阈限高度不同。 36 0 0m高度暴露 1h各项测验的综合绩效均显著下降 ,已无法满足工效要求。经过 1h的供氧 ,各项作业绩效基本恢复     

Counteracting apoptosis and necrosis with hypoxia responsive expression of Bcl-2Delta

In the encapsulated environment of biohybrid artificial organs, cells often encounter a deficiency in oxygen, which lead to apoptosis, necrosis, and lost of productivity. Two vectors with constitutive CMV promoters were constructed to examine the ability of Bcl-2Delta to help C2C12 mouse myoblasts maintain exogenous protein production under hypoxia. Two additional vectors with hypoxia-inducible promoters (5HRE) that switched on Bcl-2Delta expression based on low oxygen levels (0.0%, 0.5%, 1.0%, 2.0%, 5.0%, or 21.0%) were tested for protein productivity and protection against hypoxic stresses. A yellow fluorescent protein was used as a model protein in all vector constructs. C2C12 cells with Bcl-2Delta consistently produced more protein regardless of the oxygen level or promoter used. Cells utilizing the 5HRE rather than the CMV promoter showed an increased level of protein production as the oxygen was decreased. Among the cells with 5HRE promoters, the presence of Bcl-2Delta also increased viability and decreased apoptosis.     

Morphological characteristics of the pituitary-gonad system in high altitude hypoxia

The experiments on male rats showed that their sojourn at high altitude resulted in alterations of functional relationship between the gonadotropic pituitary cells and tests. That was characterized by synthesis inhibition and hormone release by gonadotropocytes. In the testes we found impairments in the barrier function and active substance transport in sustentocytes. Intensification of destructive processes in sustentocytes, with prolongation of adaptation duration, are mediated to a greater extent by a direct destructive high altitude factors. A decrease in hormone producing function of glandulocytes as a result of destruction processes has aggravated dysharmony of the pituitary-testes system which by the end of the second month was manifested by a marked inhibition of gonadotropic cell hormone release with preservation of the synthesis high level in them.