Modern analysis of bone loss mechanisms in microgravity

A summary of results of investigations by the author and a brief review of some literature data on human bone tissue deprived of mechanical loading (spaceflight, hypokinesia) is given. The direction and markedness of changes in bone mass--the bone mineral density and the bone mineral content--in different skeletal segments depend on their position relative to the gravity vector. A theoretically expected bone mass reduction was revealed in the trabecular structures of the bones of the lower part of the skeleton (local osteopenia). In the upper part of the skeleton, an increase in the bone mineral content is observed, which is considered as a secondary response and is due to redistribution of body fluids cephalad. The main cause of osteopenia is mechanical unloading. Arguments are presented that osteocyte osteolysis, delayed osteoblast histogenesis, and osteoclast resorption provoked by rearrangement in the hierarchy of the systems of fluid volume and ion regulation, and the endocrine control of calcium homeostasis are the main mechanisms of osteopenia.     

Multi-omics approaches to study platelet mechanisms

Platelets are small anucleate cell fragments (2–4 μm in diameter) in the blood, which play an essential role in thrombosis and hemostasis. Genetic or acquired platelet dysfunctions are linked to bleeding, increased risk of thromboembolic events and cardiovascular diseases. Advanced proteomic approaches may pave the way to a better understanding of the roles of platelets in hemostasis, and pathophysiological processes such as inflammation, metastatic spread and thrombosis. Further insights into the molecular biology of platelets are crucial to aid drug development and identify diagnostic markers of platelet activation. Platelet activation is known to be an extremely rapid process and involves multiple post-translational mechanisms at sub second time scale, including proteolysis and phosphorylation. Multi-omics technologies and biochemical approaches can be exploited to precisely probe and define these posttranslational pathways. Notably, the absence of a nucleus in platelets significantly reduces the number of present proteins, simplifying mass spectrometry-based proteomics and metabolomics approaches.     

Bone demineralization mechanisms at level of free radicals and nanoscale subsystems of bone tissue

Influence of simulated microgravity on characteristics of rat bones was investigated by electron paramagnetic resonance (EPR). For simulation of microgravity condition the hanging of animal by tail was used. The main measurements were performed for diaphysis of femoral bones. The quantity of native radicals Rn, localized in an organic matrix, and carbonate radicals CO2-, localized on a surface of bioapatite nanocrystals, were determined. The decrease of CO2- radical quantity in bones of experimental animals have shown, that due to simulation of microgravity the decrease of "collagen-nanocrystals" interaction takes place. It is shown that the EPR method open possibilities to receive the unique information about bone demineralization processes at level of free radicals and nanoscale subsystems of bone tissue.     

反义技术及其在胚胎异常发育机制研究中的应用

综述了反义研究中的几个关键问题,提出严格的ODNs设计,有效的ODNs转运和设计合理的对照等是获得良好实验结果的关键;介绍该技术在各种体外胚胎试验中的应用。     

New approaches to the study of developing wood

Two recent papers illustrate contrasting approaches to studying gene expression during development of the xylem, the tissue that transports water and solutes around higher plants. The two methods used, studying single cells differentiating in vitro and collecting samples from across the region around the cambium of poplar trees, have both revealed genes that have altered expression during xylem development.     

Daily 4-h head-up tilt is effective in preventing muscle but not bone atrophy due to simulated microgravity

To assess the potential value of intermittent artificial gravity as an efficient countermeasure, our previous studies have showed that daily 4-h standing (STD) is sufficient in counteracting muscle atrophy but not bone atrophy induced by simulated microgravity. The aim of the present study was to determine whether intermittent gravitational loading by daily 2-h or 4-h, +45 degrees head-up tilt (HUT) is more effective than STD in counteracting muscle and, particularly, bone atrophy due to simulated microgravity. Sprague-Dawley male rats weighing 290-300 g were subjected to a 28-d tail-suspension to simulate microgravity deconditioning. Daily HUT for 2, or 4 h was used to provide intermittent gravitational loading in foot-ward and tail-ward directions. The results showed that 4 h/d HUT was sufficient, and 2 h/d was less effective, in preventing adverse changes in muscle weights, fiber types, and cross-sectional areas (CSA) of muscles due to a 28-d simulated microgravity. The % protections by 4 h/d HUT in maintaining the CSAs of type I fibers in soleus, medial and lateral gastrocnemius and extensor digitorum longus muscles were 103%, 82%, 102%, and 83%, respectively. However, according to changes in physical and mechanical properties of femur, daily 4-h HUT was ineffective in attenuating the adverse changes in bone due to a 28-d simulated microgravity. Reductions in wet, dry, and ash weights and decreases in mechanical strength of femur did not show significant improvement by daily 2-h or 4-h HUT. Taken together, the findings indicate that the countermeasure effectiveness of daily 2-h or 4-h HUT for muscles is comparable with that by daily STD with the same durations. Daily 4-h HUT, as 4-h STD, is also ineffective in attenuating adverse changes in bone mass, but seems partially effective in preventing declines in mechanical properties due to simulated microgravity.     

Chemical approaches to understanding biological mechanisms

A report on work with small-molecule inhibitors of cellular processes presented at the 39th Annual Meeting of the American Society for Cell Bilogy, Washington DC, December 11-15, 1999     

New RNA-seq approaches for the study of bacterial pathogens

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Afferent mechanisms of acute responses of renal sympathetic nerve activity to microgravity

Exposure to microgravity is thought to induce an alteration of autonomic function through several afferent pathways. First, a removal of all hydrostatic gradients, which results in a large cephalad fluid shift. The fluid shift may induce an increase in transmural pressure in the cardiopulmonary region, which stimulates cardiopulmonary mechanoreceptors and baroreceptors, and elicits neurogenic responses. Secondly, an alteration of the vestibular input may modify sympathetic output via the vestibulosympathetic reflex. However, there is a lack of direct evidence for the role of cardiopulmonary mechanoreceptors, baroreceptors, and vestibular system in autonomic responses to microgravity. Accordingly, responses of renal sympathetic nerve activity (RNA) to microgravity, produced by free drop, were examined in anesthetized rats. To examine the afferent pathways of the RNA responses, the same experiment was performed in rats that had undergone labyrinthectomy, sinoaortic baroreceptor denervation (SAD), vagotomy, or SAD plus vagotomy.     

Feulgen DNA stainability of bone tumors after demineralization

Microspectrophotometric DNA analysis of archival bone tumor tissue is often impeded by previous acid demineralization, which destroys Feulgen DNA stainability. To find an alternative to acid for prospective DNA studies of bone tumors in tissue sections, Feulgen stainability of fresh osteosarcoma specimens after demineralization in neutral EDTA was investigated. The reliability of DNA analysis of weakly Feulgen-stained sections from archival tissue was also studied. Demineralization of four fresh specimens in EDTA slightly reduced Feulgen DNA stainability compared to nondemineralized preparations but did not affect the determination of ploidy level. Hydrolysis tests of one diploid and one hyperploid osteosarcoma showed that the staining relationship between control and tumor cells was not altered by EDTA pretreatment. For DNA studies of bone tumors requiring demineralization, EDTA offers a means of retaining nuclear Feulgen stainability. In 22 archival osteosarcoma specimens of varying Feulgen stainability, three different upper limits of light transmission (75, 85, and 95%) were applied to test the significance of background disturbances in relation to nuclear stain intensity. The relationship between the median total extinction of the control and tumor cell populations was not significantly affected by altering the upper transmission limit except in four poorly stained lesions. The control cells of these four specimens exhibited a median total extinction less than one-third of the maximum encountered. The results suggest that weakly stained archival specimens can be tested for selecting those appropriate for ploidy determination.     

New approaches to the study of disease in archeological New World populations.

One of the objectives of paleopathology is to clarify the role of disease in the evolution of human groups. The recovery of DNA and immunoglobulins from archeological human skeletal tissue offers a method for enhancing and expanding our knowledge about the presence and significance of disease in past human populations. DNA also might reveal the presence of genetic disease. Immunoglobulins recovered from archeological bone indicate some of the diseases to which an individual was exposed during life. This information also provides supporting evidence for anatomical observations of skeletal disease. This is illustrated by the identification of treponemal antibody in an archeological skeleton that has gross lesions suggestive of treponematosis. Similar biochemical methods could be applied to other research problems to clarify the presence of various syndromes of the inflammatory erosive arthropathies, such as rheumatoid arthritis, in New World archeological populations. Some of these syndromes are associated with DNA sequences and specific proteins that are recoverable from archeological skeletal tissue.     

Collagenase digestion of bone fragments in microgravity

The effect of a quiescent microgravity fluid environment on the activity of collagenase directed at demineralized bone fragments was investigated over a period of 10 days. Enzyme treatment resulted in greater mass loss in microgravity, with nearly three times the loss of mass during Space Shuttle mission STS-62 compared to the stationary ground control. Clinorotation enhanced the loss of mass relative to a stationary control, but this increase was still significantly less than the increase with exposure to microgravity. This suggests the detrimental influence of turbulence on the enzyme function and the benefit of using microgravity to provide both low turbulence and uniformity of unequally dense materials within the reaction chamber. The results are considered for their general applicability to a variety of bioprocessing applications that may be enhanced in microgravity. (c) 1996 John Wiley & Sons, Inc.     

微重力条件下骨矿盐丢失机理

微重力环境下骨矿盐丢失机理的研究是航天医学重要研究领域之一。骨矿盐的丢失是成骨细胞发化功能低下,成骨细胞基因表达型发生改变,使成骨细胞活性降低,导致骨持中骨钙素（ＢＧＰ）含量和碱性磷酸酶活性降低,氨基多糖含量增加,骨胶原形成异常的结果。微重力环境的下骨髓腔内脂肪含量增加,骨髓基质细胞数量减少和骨组织对骨生长因子敏感性降低是成骨细胞分化功能降低的重要原因。