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Research Containing: Mice

The impact of long-term exposure to space environment on adult mammalian organisms: a study on mouse thyroid and testis

by on 9 June 2015in Biology & Biotechnology No comment

Hormonal changes in humans during spaceflight have been demonstrated but the underlying mechanisms are still unknown. To clarify this point thyroid and testis/epididymis, both regulated by anterior pituitary gland, have been analyzed on long-term space-exposed male C57BL/10 mice, either wild type or pleiotrophin transgenic, overexpressing osteoblast stimulating factor-1. Glands were submitted to morphological and functional analysis.In thyroids, volumetric ratios between thyrocytes and colloid were measured. cAMP production in 10(-7)M and 10(-8)M thyrotropin-treated samples was studied. Thyrotropin receptor and caveolin-1 were quantitized by immunoblotting and localized by immunofluorescence. In space-exposed animals, both basal and thyrotropin-stimulated cAMP production were always higher. Also, the structure of thyroid follicles appeared more organized, while thyrotropin receptor and caveolin-1 were overexpressed. Unlike the control samples, in the space samples thyrotropin receptor and caveolin-1 were both observed at the intracellular junctions, suggesting their interaction in specific cell membrane microdomains.In testes, immunofluorescent reaction for 3beta- steroid dehydrogenase was performed and the relative expressions of hormone receptors and interleukin-1beta were quantified by RT-PCR. Epididymal sperm number was counted. In space-exposed animals, the presence of 3beta and 17beta steroid dehydrogenase was reduced. Also, the expression of androgen and follicle stimulating hormone receptors increased while lutenizing hormone receptor levels were not affected. The interleukin 1 beta expression was upregulated. The tubular architecture was altered and the sperm cell number was significantly reduced in spaceflight mouse epididymis (approx. -90% vs. laboratory and ground controls), indicating that the space environment may lead to degenerative changes in seminiferous tubules.Space-induced changes of structure and function of thyroid and testis/epididymis could be responsible for variations of hormone levels in human during space missions. More research, hopefully a reflight of MDS, would be needed to establish whether the space environment acts directly on the peripheral glands or induces changes in the hypotalamus-pituitary-glandular axis.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/22558148

Microgravity alters the expression of salivary proteins

by on 9 June 2015in Biology & Biotechnology No comment

Spaceflight provides a unique opportunity to study how physiologic responses are influenced by the external environment. Microgravity has been shown to alter the function of a number of tissues and organ systems. Very little, however, is known about how microgravity affects the oral cavity. The rodent model is useful for study in that their salivary gland morphology and physiology is similar to that of humans. Useful also is the fact that saliva, a product of the salivary glands with a major role in maintaining oral health, can be easily collected in humans whereas the glands can be studied in experimental animals. Our working hypothesis is that expression of secretory proteins in saliva will respond to microgravity and will be indicative of the nature of physiologic reactions to travel in space. This study was designed to determine which components of the salivary proteome are altered in mice flown on the US space shuttle missions and to determine if a subset with predictive value can be identified using microscopy and biochemistry methods. The results showed that the expression of secretory proteins associated with beta-adrenergic hormone regulated responses and mediated via the cyclic AMP pathway was significantly altered, whereas that of a number of unrelated proteins was not. The findings are potentially applicable to designing a biochemical test system whereby specific salivary proteins can be biomarkers for stress associated with travel in space and eventually for monitoring responses to conditions on earth.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/24984624

Simulated microgravity promoted differentiation of bipotential murine oval liver stem cells by modulating BMP4/Notch1 signaling

by on 9 June 2015in Biology & Biotechnology No comment

Faster growth and differentiation of liver stem cells to hepatocyte is one of the key factors during liver regeneration. In recent years, simulated microgravity, a physical force has shown to differentially regulate the differentiation and proliferation of stem cells. In the present work, we studied the effect of simulated microgravity on differentiation and proliferation of liver stem cells. The cells were subjected to microgravity, which was simulated using indigenously fabricated 3D clinostat. Proliferation, apoptosis, immunofluorescence assays and Western blot analysis were carried out to study the effects of simulated microgravity on liver stem cells. Microgravity treatment for 2 h enhanced proliferation of stem cells by twofold without inducing apoptosis and compromising cell viability. Analysis of hepatocyte nuclear factor 4-alpha (HNF4-alpha) expression after 2 h of microgravity treatment revealed that microgravity alone can induce the differentiation of stem cells within 2-3 days. Probing bone morphogenic protein 4 (BMP4) and Notch1 in microgravity treated stem cells elaborated downregulation of Notch1 and upregulation of BMP4 after 2 days of incubation. Further, blocking BMP4 using dorsomorphin and chordin conditioned media from chordin plasmid transfected cells attenuated microgravity mediated differentiation of liver stem cells. In conclusion, microgravity interplays with BMP4/Notch1 signaling in stem cells thus inducing differentiation of stem cells to hepatocytes. Present findings can be implicated in clinical studies where microgravity activated stem cells can regenerate the liver efficiently after liver injury.

Related URLs:
http://ovidsp.ovid.com/ovidweb.cgi?T=JS&CSC=Y&NEWS=N&PAGE=fulltext&D=emed10&AN=2011438430
http://sfxhosted.exlibrisgroup.com/mayo?sid=OVID:embase&id=pmid:&id=doi:10.1002%2Fjcb.23110&issn=0730-2312&isbn=&volume=112&issue=7&spage=1898&pages=1898-1908&date=2011&title=Journal+of+Cellular+Biochemistry&atitle=Simulated+microgravity+promoted+differentiation+of+bipotential+murine+oval+liver+stem+cells+by+modulating+BMP4%2FNotch1+signaling&aulast=Majumder&pid=%3Cauthor%3EMajumder+S.%3C%2Fauthor%3E&%3CAN%3E2011438430%3C%2FAN%3E

Neural precursor cells form rudimentary tissue-like structures in a rotating-wall vessel bioreactor

by on 9 June 2015in Biology & Biotechnology No comment

We have analyzed the biology of embryonic, epidermal growth factor-responsive murine neural precursor cells cultured in the high-aspect ratio vessel (HARV). Within 2-3 d of rotary-cell culture, such cells formed multiple, macroscopic, three-dimensional structures that were orders of magnitude larger than the cellular clusters ("neurospheres") formed by these cells in conventional stationary-flask cultures. Each HARV structure was composed of a multilayered cellular shell surrounding one or more central cavities that were bordered by pyknotic cell nuclei. Although the cells in the HARV structures were more pleomorphic than those in neurospheres, the structures did not appear to represent primitive neural tumors: the formation of HARV structures by precursor cells was not an irreversible phenotypic change, and the structures dill not originate front the clonal expansion of single-progenitor cells; the growth rate and invasiveness of the cells in HARVs were less than those in flasks; and HARV-cultured cells did not form tumors after subcutaneous inoculation into the Hanks of NOD-scid/scid mice. Immunohistochemical analysis suggested that HARV structures might be novel "proto-tissues" characterized by a crude. but organized, architecture, with a surface laver of immature proliferating cells (nestin- and proliferating cell nuclear antigen-positive) that enclosed strata of more differentiated cells (beta -tubulin III- and glial fibrillary acidic protein-positive) within. Rotary-cell culture may have significant implications for the eventual utility. of neural precursors for clinical neurotransplantation.

Related URLs:
<Go to ISI>://WOS:000168690900004

The effect of spaceflight on mouse olfactory bulb volume, neurogenesis, and cell death indicates the protective effect of novel environment

by on 9 June 2015in Biology & Biotechnology No comment

Space missions necessitate physiological and psychological adaptations to environmental factors not present on Earth, some of which present significant risks for the central nervous system (CNS) of crewmembers. One CNS region of interest is the adult olfactory bulb (OB), as OB structure and function are sensitive to environmental- and experience-induced regulation. It is currently unknown how the OB is altered by spaceflight. In this study, we evaluated OB volume and neurogenesis in mice shortly after a 13-day flight on Space Shuttle Atlantis [Space Transport System (STS)-135] relative to two groups of control mice maintained on Earth. Mice housed on Earth in animal enclosure modules that mimicked the conditions onboard STS-135 (AEM-Ground mice) had greater OB volume relative to mice maintained in standard housing on Earth (Vivarium mice), particularly in the granule (GCL) and glomerular (GL) cell layers. AEM-Ground mice also had more OB neuroblasts and fewer apoptotic cells relative to Vivarium mice. However, the AEM-induced increase in OB volume and neurogenesis was not seen in STS-135 mice (AEM-Flight mice), suggesting that spaceflight may have negated the positive effects of the AEM. In fact, when OB volume of AEM-Flight mice was considered, there was a greater density of apoptotic cells relative to AEM-Ground mice. Our findings suggest that factors present during spaceflight have opposing effects on OB size and neurogenesis, and provide insight into potential strategies to preserve OB structure and function during future space missions.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/24744382

Ultrasound Guided Combined Cryoablation and Microencapsulated 5-Fluorouracil Inhibits Growth of Human Prostate Tumors in Xenogenic Mouse Model Assessed by Luminescence Imaging

by on 9 June 2015in Biology & Biotechnology No comment

Modern approaches to minimally invasive ablative treatment of solid tumors involve the use of miniature instruments and combined treatments. These can be enhanced with ultrasound imaging that depicts tumor margins; facilitates guidance, delivery, and dosage of local chemotherapy; and can monitor the effectiveness of the treatment. This paper describes the advantages of ultrasound guided cryosurgery combined with local chemotherapy delivered in multilamellar, echogenic microcapsules of 5-FU (“μcaps”) using a xenograft tumor model. Genetically engineered bioluminescent human prostate tumor cells, DU-145Luc+, were implanted subcutaneously into athymic nude mice. Experiments were designed to mimic the situation where palliative cryoablation spares a portion of the tumor so that the combined effect of cryosurgery and focal injections of chemotherapeutic microcapsules could be evaluated. Eighteen (18) tumors were treated with percutaneous partial cryoablation or interstitial chemoablation, or a combination of both. A single F/T cycle was applied to tumor and micro-encapsulated chemotherapy is delivered at outer margin of frozen tumor in two opposite sites. Results show that the tumor and cryosurgical kill zone contours were seen with both the bio-luminescence assay (BLI) and ultrasonography (US). US can easily detect as little as 2 μl of echogenic μcaps, and monitor their lifetime in the tumor tissue. BLI was determinant in showing that minute amounts of microcapsule chemotherapy (38.7 ng of 5-FU/g tumor) dramatically inhibited tumor growth starting within two days after injection. The mean BLI emitted by control tumors was 5.6 times greater at Day 4 than the BLI measurements from tumors treated with 5-FU μcaps (p=0.036). By Day 7, BLI values from the control tumors were still 2.7 times greater than those treated with 5-FU μcaps (p<0.01). In tumors treated by partial cryoablation, the mean BLI of viable tumor cells was 20 times less at day 3 (p=0.05) and 46% less at day 7 than the non-treated tumors. The combined treatment produced a dramatic inhibition of tumor growth that lasted throughout the 7-day study. The BLI measured from viable tumor cells in non-treated tumors was 34 times greater at day 3 and more than 350 times greater at day 7 than those treated by combined cryoablation and 5-FU μcaps. The results demonstrated, for the first time, that a single moderate freeze of a human prostate tumor combined with bi-focal peripheral microcapsule chemotherapy (5-FU) has a better and longer inhibitory effect on tumor growth compared to the growth inhibition rendered by cryosurgery or local microcapsule chemo-therapy alone. This shows promise for a new, focal, combined ablative modality using US guided deposition of microencapsulated drug(s) and echogenic markers deposited in the hypothermic margin of tumors which could enhance the efficacy of cryoablation of prostate cancers.

Related URLs:
http://tct.sagepub.com/content/3/2/135.abstract

Percutaneous tumor ablation: microencapsulated echo-guided interstitial chemotherapy combined with cryosurgery increases necrosis in prostate cancer

by on 9 June 2015in Biology & Biotechnology No comment

This study aimed at confirming the increased growth inhibition (GI) of human prostate tumors produced by a intentionally palliative combination treatment of cryochemotherapy, i.e., partial cryoablation (CA) followed by intratumor partial chemotherapy with injection of microencapsulated 5-fluorouracil (MCC/5FU) at the ice ball (IB) periphery. We report the local effectiveness of cryochemotherapy compared to chemotherapy only with using multiple injections of MCC/5FU spaced out to maximize cumulative effect of sustained release of 5-fluorouracil (5FU) during a 21-day period. Prostate bioluminescent tumor cells – DU145 Luc+ – were implanted sub-cutaneously and bilaterally in each flank of nude mice. Tumors were treated with: (i) cryoablation alone (CA), causing necrosis in approximately 45% of the tumor volume; (ii) cryo-chemotherapy (CA+MCC/5FU), a combined regimen consisting of partial CA followed immediately and on day 14 by ultrasound assisted, intra-tumor injections (40 mul) of MCC/5FU( 0.81 ng/mm3 of tumor) containing Ethiodol (IPO) an imaging contrast agent, on two opposite sides of the unfrozen part of tumor; (iii) intratumor chemotherapy (MCC/5FU), consisting of three successive intra-tumor injections of microencapsulated 5FU on two opposite sides on Day 0, 4, and 11, and (iv) control series (MM), consisting of a single injection of echogenic microcapsules (mucaps) containing IPO but no 5FU. Tumor growth and viability were followed during a 21-day period with using biometric measurements, bioluminescent imaging (BLI) and ultrasonography (US), and then animals were sacrificed. CA, spared 54.4% of the tumor volume and the IB kill ratio was 0.4 +/-0.9. The maximum tumor volume reduction observed by Day 3 was short-lived as re-growth became significant by Day 6. CA+ MCC/5FU spared 55.6% of the tumor volume and the IB kill ratio was 0.54 +/- 0.12. The viable tumor cells, as measured by BLI remained at preoperative levels. After 11 days CA+ MCC/5FU limited the growth of the partially ablated tumors to only 10.6% of the growth of CA treated tumors (p=0.04). By Day 18 the CA+MCC/5FU had inhibited tumor growth by 78% compared to the CA treated tumors (p=0.05) and after 21 days the growth was inhibited by 71% (p=0.04) compared to more than 650% growth in the MM group and 600% growth in the CA treated group. The two injections of MCC/5FU produced a visible focal necrosis in 55% of the tumors. MCC/5FU proved effective by themselves and reduced the growth of prostate tumor volumes by 51% (p=0.025) compared to MM controls during the 21 days. Focal necrosis was macroscopically visible at the site of 66% of the tumors injected only with MCC/5FU. The BLI clearly showed zones of reduced tumor cell viability at the injection sites. The mean number of bioluminescent (viable) tumor cells, remained below preoperative levels for the first 6 days and then increased at a rate approximately 20% that of the growth of control tumor cells. The chemoablative effects of intentionally limited doses of MCC/5FU injected within the IB margin augment the effects of incomplete cryoablation in this prostate tumor model, with dramatic tumor GI and directionally increased necrosis dimensions compared to CA alone, confirming the results of a previous study. Our results indicate the potential advantages of our combination cryochemotherapy that utilizes different mechanisms to kill tumor cells and retard tumor growth in the region surrounding the IB where tumor cells escape the lethal effects of cryosurgery. The study suggests that cryochemotherapy may become a more predictable technique that could be indicated as an adjuvant or an alternative to palliative therapy of hormone refractory prostate cancer (HRPC).

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/19445538

Microarray analysis of spaceflown murine thymus tissue reveals changes in gene expression regulating stress and glucocorticoid receptors

by on 9 June 2015in Biology & Biotechnology No comment

The detrimental effects of spaceflight and simulated microgravity on the immune system have been extensively documented. We report here microarray gene expression analysis, in concert with quantitative RT-PCR, in young adult C57BL/6NTac mice at 8 weeks of age after exposure to spaceflight aboard the space shuttle (STS-118) for a period of 13 days. Upon conclusion of the mission, thymus lobes were extracted from space flown mice (FLT) as well as age- and sex-matched ground control mice similarly housed in animal enclosure modules (AEM). mRNA was extracted and an automated array analysis for gene expression was performed. Examination of the microarray data revealed 970 individual probes that had a 1.5-fold or greater change. When these data were averaged (n = 4), we identified 12 genes that were significantly up- or down-regulated by at least 1.5-fold after spaceflight (P < or = 0.05). The genes that significantly differed from the AEM controls and that were also confirmed via QRT-PCR were as follows: Rbm3 (up-regulated) and Hsph110, Hsp90aa1, Cxcl10, Stip1, Fkbp4 (down-regulated). QRT-PCR confirmed the microarray results and demonstrated additional gene expression alteration in other T cell related genes, including: Ctla-4, IFN-alpha2a (up-regulated) and CD44 (down-regulated). Together, these data demonstrate that spaceflight induces significant changes in the thymic mRNA expression of genes that regulate stress, glucocorticoid receptor metabolism, and T cell signaling activity. These data explain, in part, the reported systemic compromise of the immune system after exposure to the microgravity of space.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/20213684

Effects of simulated microgravity on mammalian fertilization and preimplantation embryonic development in vitro

by on 9 June 2015in Biology & Biotechnology No comment

Objective: To study the effects of simulated microgravity on mammalian fertilization and preimplantation embryonic development in vitro with the use of a horizontal clinostat device. Design: Controlled animal study. Setting: Research laboratory at a university medical school. Animal(s): B6D2F1 (C57BL/6 x DBA/2) and ICR mice between 8 and 10 weeks old. Intervention(s): The first experiment was performed to investigate whether gravity is required for fertilization in vitro under three conditions: clinostat rotation, rotational control, and stationary control. In the second experiment, one-cell embryos were cultured under each condition and their morphology and viability were assessed at 96 hours. Main Outcome Measure(s): The fertilized numbers and embryonic numbers at the morula and blastocyst stages were recorded in each condition. Result(s): In the first experiment, there were no statistically significant differences in the efficiency of achieving normal fertilization in vitro among the conditions. In the second experiment, there was a statistically significant decrease in the number of embryos reaching the morula and blastocyst stages after 96 hours in culture under clinostat rotation. Conclusion(s): These results suggest that the process of fertilization in vitro is not sensitive to the gravitational vector. However, the possibility exists that the frequency of early embryonic lethality is increased by microgravity. (Fertil Steril(R) 2000;74:1142-7. (C) 2000 by American Society for Reproductive Medicine).

Related URLs:
<Go to ISI>://WOS:000165897300014