scholarly journals Effects of deuterium oxide on cell growth and vesicle speed in RBL-2H3 cells

Author(s):  
Roshni S Kalkur ◽  
Andrew C Ballast ◽  
Ashley R Triplett ◽  
Kathrin Spendier

For the first time we show the effects of deuterium oxide on the cell growth and vesicle transport in rat basophilic leukemia (RBL-2H3) cells. RBL-2H3 cells cultured with 15 moles/L deuterium showed decreased cell growth which was attributed to cells not doubling their DNA content. Experimental observations also showed an increase in vesicle speed for cells cultured in deuterium oxide. This increase in vesicle speed was not observed in deuterium oxide cultures treated with a microtubule-destabilizing drug suggesting that deuterium oxide affects microtubule-dependent vesicle transport.

2014 ◽  
Author(s):  
Roshni S Kalkur ◽  
Andrew C Ballast ◽  
Ashley R Triplett ◽  
Kathrin Spendier

For the first time we show the effects of deuterium oxide on the cell growth and vesicle transport in rat basophilic leukemia (RBL-2H3) cells. RBL-2H3 cells cultured with 15 moles/L deuterium showed decreased cell growth which was attributed to cells not doubling their DNA content. Experimental observations also showed an increase in vesicle speed for cells cultured in deuterium oxide. This increase in vesicle speed was not observed in deuterium oxide cultures treated with a microtubule-destabilizing drug suggesting that deuterium oxide affects microtubule-dependent vesicle transport.


2014 ◽  
Author(s):  
Roshni S Kalkur ◽  
Andrew Ballast ◽  
Ashley R Triplett ◽  
Kathrin Spendier

For the first time we show the effects of deuterium oxide on the cell cycle and vesicle transport in rat basophilic leukemia (RBL-2H3) cells. RBL-2H3 cultured with 15 moles/L deuterium oxide were observed to halt near the G2-M-phase of the cell cycle. Experimental observations also showed an increase in vesicle speed. This increase in vesicle speed was not observed in cultures treated with a microtubule-destabilizing drug suggesting that deuterium oxide affects microtubule-dependent vesicle transport.


2021 ◽  
Vol 87 (6) ◽  
Author(s):  
Katsuya Fuchino ◽  
Helena Chan ◽  
Ling Chin Hwang ◽  
Per Bruheim

ABSTRACT The alphaproteobacterium Zymomonas mobilis exhibits extreme ethanologenic physiology, making this species a promising biofuel producer. Numerous studies have investigated its biology relevant to industrial applications and mostly at the population level. However, the organization of single cells in this industrially important polyploid species has been largely uncharacterized. In the present study, we characterized basic cellular behavior of Z. mobilis strain Zm6 under anaerobic conditions at the single-cell level. We observed that growing Z. mobilis cells often divided at a nonmidcell position, which contributed to variant cell size at birth. However, the cell size variance was regulated by a modulation of cell cycle span, mediated by a correlation of bacterial tubulin homologue FtsZ ring accumulation with cell growth. The Z. mobilis culture also exhibited heterogeneous cellular DNA content among individual cells, which might have been caused by asynchronous replication of chromosome that was not coordinated with cell growth. Furthermore, slightly angled divisions might have resulted in temporary curvatures of attached Z. mobilis cells. Overall, the present study uncovers a novel bacterial cell organization in Z. mobilis. IMPORTANCE With increasing environmental concerns about the use of fossil fuels, development of a sustainable biofuel production platform has been attracting significant public attention. Ethanologenic Z. mobilis species are endowed with an efficient ethanol fermentation capacity that surpasses, in several respects, that of baker’s yeast (Saccharomyces cerevisiae), the most-used microorganism for ethanol production. For development of a Z. mobilis culture-based biorefinery, an investigation of its uncharacterized cell biology is important, because bacterial cellular organization and metabolism are closely associated with each other in a single cell compartment. In addition, the current work demonstrates that the polyploid bacterium Z. mobilis exhibits a distinctive mode of bacterial cell organization, likely reflecting its unique metabolism that does not prioritize incorporation of nutrients for cell growth. Thus, another significant result of this work is to advance our general understanding in the diversity of bacterial cell architecture.


2021 ◽  
Author(s):  
Hyunbum Jang ◽  
Iris Nira Smith ◽  
Charis Eng ◽  
Ruth Nussinov

AbstractTumor suppressor PTEN dephosphorylates signaling lipid PIP3 produced by PI3Ks. Abundant PIP3 promotes cell growth and proliferation. PTEN is the second most highly mutated protein in cancer and is drugless. The detailed mechanism at the membrane of this pivotal phosphatase is unknown hindering understanding and drug discovery. Here for the first time, exploiting explicit solvent simulations, we tracked full-length PTEN trafficking from the cytosol to the membrane, its interaction with membranes composed of zwitterionic phosphatidylcholine and anionic phosphatidylserine and phosphatidylinositol, including signaling lipids PIP2 and PIP3, and moving away from the zwitterionic and getting absorbed onto the anionic membrane that harbors the PIP3. PIP3 then allosterically unfolds the N-terminal PIP2 binding domain, translocating it to the membrane where its polybasic motif interacts with PIP2, localizing on microdomains enriched in signaling lipids, as PI3K does. Finally, we determined PTEN catalytic action at the membrane, all in line with available experimental observations.


2016 ◽  
Vol 473 (8) ◽  
pp. 1037-1046 ◽  
Author(s):  
Bo Zhang ◽  
Xiao-Jian Hu ◽  
Xiao-Qiang Wang ◽  
Jean-François Thériault ◽  
Dao-Wei Zhu ◽  
...  

Human 3α-HSD3 (3α-hydroxysteroid dehydrogenase type 3) plays an essential role in the inactivation of the most potent androgen 5α-DHT (5α-dihydrotestosterone). The present study attempts to obtain the important structure of 3α-HSD3 in complex with 5α-DHT and to investigate the role of 3α-HSD3 in breast cancer cells. We report the crystal structure of human 3α-HSD3·NADP+·A-dione (5α-androstane-3,17-dione)/epi-ADT (epiandrosterone) complex, which was obtained by co-crystallization with 5α-DHT in the presence of NADP+. Although 5α-DHT was introduced during the crystallization, oxidoreduction of 5α-DHT occurred. The locations of A-dione and epi-ADT were identified in the steroid-binding sites of two 3α-HSD3 molecules per crystal asymmetric unit. An overlay showed that A-dione and epi-ADT were oriented upside-down and flipped relative to each other, providing structural clues for 5α-DHT reverse binding in the enzyme with the generation of different products. Moreover, we report the crystal structure of the 3α-HSD3·NADP+·4-dione (4-androstene-3,17-dione) complex. When a specific siRNA (100 nM) was used to suppress 3α-HSD3 expression without interfering with 3α-HSD4, which shares a highly homologous active site, the 5α-DHT concentration increased, whereas MCF7 cell growth was suppressed. The present study provides structural clues for 5α-DHT reverse binding within 3α-HSD3, and demonstrates for the first time that down-regulation of 3α-HSD3 decreases MCF7 breast cancer cell growth.


2010 ◽  
Vol 129-131 ◽  
pp. 1029-1033
Author(s):  
Xiang Dong Ma ◽  
Xiao Ming Wu ◽  
Hai Xia Duan ◽  
Xing Ma ◽  
Tao Fu

Nanosized β-tricalcium phosphate (TCP) material was produced in this study using a wet precipitation method and characterized by transmission electron microscopy (TEM) and X-ray diffraction (XRD). Human ovarian sarcoma SKOV-3 cells were cultured and the influence of nanoscale β-TCP particles on SKOV-3 cell behavior was studied in vitro. As a result, β-TCP nanoparticles with average size of 100 nm were obtained. Cell growth of SKOV-3 cells was noticeably declined in the presence of β-TCP nanoparticles (200ng/ml). The distribution of cell cycle for SKOV-3 cells cultured with and without β-TCP nanomaterials was quite different. In G1 phase of cell cycle, the percentage of SKOV-3 cells cultured in the absence of β-TCP nanoparticles was significantly lower than that cultured in the presence of β-TCP nanoparticles (p<0.01). In S phase of cell cycle, on the other hand, the percentage of SKOV-3 cells cultured without β-TCP nanoparticles was noticeably increased compared with that cultured with β-TCP nanoparticles (p<0.01). Moreover, the expression of proliferating cell nuclear antigen (PCNA) in SKOV-3 cells cultured in medium containing 200ng/ml β-TCP nanopaticles was significantly lower than that in the cells cultured without β-TCP nanoparticles (p<0.01). In conclusion, the nanoscale β-TCP material synthesized in this study can exert anti-tumor effects on SKOV-3 cells through mechanisms of cell growth inhibition, downregulation of PCNA expression and cell cycle arrest at G1 phase.


2016 ◽  
pp. 121-131 ◽  
Author(s):  
E. FILOVÁ ◽  
B. JAKUBCOVÁ ◽  
I. DANILOVÁ ◽  
E. KUŽELOVÁ KOŠŤÁKOVÁ ◽  
T. JAROŠÍKOVÁ ◽  
...  

For biodegradable porous scaffolds to have a potential application in cartilage regeneration, they should enable cell growth and differentiation and should have adequate mechanical properties. In this study, our aim was to prepare biocompatible scaffolds with improved biomechanical properties. To this end, we have developed foam scaffolds from poly-Ɛ-caprolactone (PCL) with incorporated chitosan microparticles. The scaffolds were prepared by a salt leaching technique from either 10 or 15 wt% PCL solutions containing 0, 10 and 20 wt% chitosan microparticles, where the same amount and size of NaCl was used as a porogen in all the cases. PCL scaffolds without and with low amounts of chitosan (0 and 10 wt% chitosan) showed higher DNA content than scaffolds with high amounts of chitosan during a 22-day experiment. 10 wt% PCL with 10 and 20 wt% chitosan showed significantly increased viscoelastic properties compared to 15 wt% PCL scaffolds with 0 and 10 wt% chitosan. Thus, 10 wt% PCL scaffolds with 0 wt% and 10 wt% chitosan are potential scaffolds for cartilage regeneration.


Author(s):  
С.В. Смирнова ◽  
Т.Н. Шапиро ◽  
Е.В. Игонина ◽  
С.К. Абилев

Изучали генотоксическое действие бактерицидных средств диоксидина, фурацилина и налидиксовой кислоты на клетки дейтерированной культуры lux-биосенсора E.coli MG1655 (pColD::lux), люминесцирующего в результате активации промотора гена колицина colD в ответ на повреждение ДНК. Впервые показано, что оксид дейтерия (D2O) в концентрации 9% усиливает SOS-ответ, индуцированный исследуемыми лекарственными препаратами, в 1,6-2,8 раза в клетках E. coli. We studied the genotoxic effect of bactericidal agents: dioxine, furaciline and nalidixic acid on cells of the deuterated culture lux-biosensor E. coli MG1655 (pColD::lux), which luminesces as a result of activation of the colicin gene promoter colD in response to DNA damage. For the first time, it was shown that deuterium oxide (D2O) at a concentration of 9% increases the SOS response by 1.6-2.8 times in E. coli cells induced by the studied drugs.


Genetics ◽  
1994 ◽  
Vol 136 (2) ◽  
pp. 465-474 ◽  
Author(s):  
A D Hartley ◽  
M P Ward ◽  
S Garrett

Abstract The growth defect associated with the loss of yeast A kinase activity can be alleviated by the overexpression or deletion of two other kinases, Sch9 and Yak1, respectively. Using tests of epistasis, we have shown that Sch9 and Yak1 define separate signaling pathways and must, therefore, suppress the A kinase defect by different mechanisms. Nevertheless, the Yak1 kinase appears to regulate cellular processes that are under A kinase control. For example, acquisition of heat resistance is correlated with Yak1 kinase activity, such that YAK1-overexpressing cells are over 200-fold more resistant than isogenic yak1 strains. These results, for the first time, associate a phenotype, other than suppression of the A kinase growth defect, with the loss of Yak1 activity and argue a broader role for the Yak1 kinase in cell growth.


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