scholarly journals Elimination of [PSI+] Prion and Influence on Yeast Cell Growth

Author(s):  
Justina Versockienė ◽  
Neda Jonutytė-Trembo ◽  
Vitalij Novickij ◽  
Eglė Lastauskienė

Abstract Background Prions are proteinaceous infectious particles that act as pathogens and cause the development of lethal neurodegenerative diseases in humans and other animals. Yeast Saccharomyces cerevisiae is a widespread model system in which mechanisms of prion induction and elimination have been identified. New and safe substances and methods are being sought to cure cells of prion proteins. It is particularly important that by treating cells from prions and restoring them from the [PSI+] to the [psi−] form, the primary growth of the cells is restored. One of the main objectives of this study was to determine the growth dynamics of S. cerevisiae cells with different [PSI+] prion variants, cells that have lost [PSI+] prion variants, and cells that never had [PSI+] prion variants. Results In this research, we applied GuHCl and combined GuHCl and PEF treatment against [PSI+] prion. We evaluated cells culture growth dynamics – optical density and doubling time and determined that method of [PSI+] prion elimination does not affect cell doubling time. Also, we found that both elimination methods affect the optical density reached by [psi−] cells. However, the cells in which the [PSI+] prion has been eliminated by GuHCl alone are able to reach the same optical density as unaffected [psi−] cells and higher optical density than the affected [psi−] cells by GuHCl alone. Conclusions These findings indicate the potential long-term positive effect of [PSI+] prion on cell growth, which persists after [PSI+] removal.

1997 ◽  
Vol 272 (2) ◽  
pp. F273-F282 ◽  
Author(s):  
T. B. Kinane ◽  
I. Kang ◽  
A. Chu ◽  
S. H. Chin ◽  
L. Ercolani

The protooncogene G alpha(i-2) plays a pivotal role in signaling pathways that control renal cell growth and differentiation. Mitogen-activated protein kinases (MAPKs) are potential downstream effectors for G alpha(i-2) in these pathways. In predifferentiated LLC-PK1 renal cells, the temporal maximal expression of G alpha(i-2) coincided with maximal activation of MAPK(p42/p44). By contrast, pertussis toxin treatment of these cells inhibited cell growth and reduced MAPK(p42/p44) activity by 30%. These findings reflected upstream activation of MAPK kinase (MEK1), as transient transfection of cells with a plasmid encoding a constitutively active form of MEK1 increased MAPK(p42/p44) activity and cell growth, whereas treatment with PD-098059, an inhibitor of MEK1 activity, reduced MAPK(p42/p44) activity and cell growth. Expression of a guanosinetriphosphatase (GTPase)-deficient G alpha(i-2) in these cells increased MAPK(p42/p44) activity and correspondingly reduced cell doubling time from 24 to 10 h without altering the activity of Raf-1 or c-Jun/stress-activated protein kinases (SAPKs). By contrast, expression of a GTPase-deficient G alpha(i-3) in these cells reduced both their cell doubling time by 30% and MAPK(p42/p44) activity by 60%. As the known MEKK isoforms (MEKK1, -2, and -3) can also activate SAPKs, these findings suggest the GTP-charged G alpha(i-2) subunit transduces growth signals in renal cells via activation of MAPK(p42/p44) and that such activation may be linked to pathways containing novel MEKK isoforms that preferentially activate MEKs.


2008 ◽  
Vol 51 (3) ◽  
pp. 165-172 ◽  
Author(s):  
Lenka Brůčková ◽  
Tomáš Soukup ◽  
Jiří Moos ◽  
Martina Moosová ◽  
Jana Pavelková ◽  
...  

The major functions of granulosa cells (GCs) include the production of steroids, as well as a myriad of growth factors to interact with the oocyte during its development within the ovarian follicle. Also FSH stimulates GCs to convert androgens (coming from the thecal cells) to estradiol by aromatase. However, after ovulation the GCs produce progesterone that may maintain a potential pregnancy. Experiments with human GCs are mainly focused on the purification of GCs from ovarian follicular fluid followed by FACS analysis or short-term cultivation. The aim of our study was to cultivate GCs for a long period, to characterize their morphology and phenotype. Moreover, we have cultivated GCs under gonadotropin stimulation in order to simulate different pathological mechanisms during folliculogenesis (e.g. ovarian hyperstimulation syndrome). GCs were harvested from women undergoing in vitro fertilization. Complex oocyte-cumulus oophorus was dissociated by hyaluronidase. The best condition for transport of GCs was optimized as short transport in follicular fluid at 37 °C. GCs expansion medium consisted of DMEM/F12, 2 % FCS, ascorbic acid, dexamethasone, L-glutamine, gentamycine, penicillin, streptomycin and growth factors (EGF, bFGF). GCs transported in follicular fluid and cultivated in 2 % FCS containing DMEM/F12 medium supplemented with follicular fluid presented increased adhesion, proliferation, viability and decreased doubling time. Cell viability was 92 % and mean cell doubling time was 52 hrs. We have optimized transport and cultivation protocols for long-term cultivation of GCs.


1988 ◽  
Vol 43 (1-3) ◽  
pp. 151-173 ◽  
Author(s):  
Samuel M. Cohen ◽  
Leon B. Ellwein

Author(s):  
Valentina Yurievna Stupko ◽  
Natalya Vasilevna Zobova ◽  
Yuri Leonidovich Gurevich

High biological activity of nanoparticles (NPs) makes them promising for using in fertilizers composition. Design of such products is impossible without detection of active substance influence mechanisms on growth and development of plant as well as without detection of critical points, when influence of the investigated agent is at its maximum. Biogenic ferrihydrites (FH) are promising regulators of plant cell antioxidant system activity, also under edaphic stresses. Callus cultures are well known as convenient model system for screening of tissue and cell response to different environmental exposure. Investigation results of FH NP`s, which were produced by authors, influence on growth and development of callus culture of soft spring wheat mature wheat embryos of are presented in present article. The absence of competitive in relation to 2,4-D influence of involved levels of FH NPs (1 and 10 mg/l) on dedifferentiation process of embryo tissues was indicated. The presence of investigated agent induced the decrease of culture growth rate. However, postponed positive effect manifested in reduction of ageing rate after passing to fresh medium, the bigger, the higher had been FH NP`s level at the previous cultivation stage. The present effect may be of critical importance under field conditions under the influence of unfavorable edaphic and climatic factors, which adds further credence the potential of presented NPs applicability in agriculture.


2021 ◽  
Vol 11 (8) ◽  
pp. 1025
Author(s):  
Miodrag Dragoj ◽  
Jasmina Stojkovska ◽  
Tijana Stanković ◽  
Jelena Dinić ◽  
Ana Podolski-Renić ◽  
...  

Background: Various three-dimensional (3D) glioblastoma cell culture models have a limited duration of viability. Our aim was to develop a long-term 3D glioblastoma model, which is necessary for reliable drug response studies. Methods: Human U87 glioblastoma cells were cultured in alginate microfibers for 28 days. Cell growth, viability, morphology, and aggregation in 3D culture were monitored by fluorescent and confocal microscopy upon calcein-AM/propidium iodide (CAM/PI) staining every seven days. The glioblastoma 3D model was validated using temozolomide (TMZ) treatments 3 days in a row with a recovery period. Cell viability by MTT and resistance-related gene expression (MGMT and ABCB1) by qPCR were assessed after 28 days. The same TMZ treatment schedule was applied in 2D U87 cell culture for comparison purposes. Results: Within a long-term 3D model system in alginate fibers, U87 cells remained viable for up to 28 days. On day 7, cells formed visible aggregates oriented to the microfiber periphery. TMZ treatment reduced cell growth but increased drug resistance-related gene expression. The latter effect was more pronounced in 3D compared to 2D cell culture. Conclusion: Herein, we described a long-term glioblastoma 3D model system that could be particularly helpful for drug testing and treatment optimization.


Author(s):  
Ol'ga Gladysheva ◽  
Oksana Artyuhova ◽  
Vera Svirina

The results of long-term research in experiments with crop rotations with different clover saturation are presented. It is shown that the cluster has a positive effect on the main indicators of vegetation of dark-gray forest soil. The introduction of two fields of perennial grasses into the six-field crop rotation significantly increases both the humus reserves and increases the productivity of arable land by 1.5–2 times compared to the crop rotation with a field of pure steam.


1985 ◽  
Vol 110 (3) ◽  
pp. 329-337 ◽  
Author(s):  
G. A. Schuiling ◽  
H. Moes ◽  
T. R. Koiter

Abstract. The effect of pretreatment in vivo with oestradiol benzoate on in vitro secretion of LH and FSH was studied in long-term ovariectomized (OVX) rats both at the end of a 5-day continuous in vivo pretreatment with LRH and 4-days after cessation of such LRH pretreatment. Rats were on day 0 sc implanted with osmotic minipumps which released LRH at the rate of 250 ng/h. Control rats were implanted with a piece of silicone elastomer with the dimensions of a minipump. On days 2 and 4 the rats were injected with either 3 μg EB or with oil. On day 5 part of the rats were decapitated and the in vitro autonomous (i.e. non-LRH-stimulated) and 'supra-maximally' LRHstimulated release of LH and FSH was studied using a perifusion system. From other rats the minipumps were removed on day 5 and perifusion was performed on day 9. On the 5th day of the in vivo LRH pretreatment the pituitary LH/FSH stores were partially depleted; the pituitaries of the EB-treated rats more so than those of the oil-injected rats. EB alone had no significant effect on the content of the pituitary LH- and FSH stores. On day 9, i.e. 4 days after removal of the minipumps, the pituitary LH and FSH contents had increased in both the oil- and the EB injected rats, but had not yet recovered to control values. In rats not subjected to the 5-days pretreatment with LRH EB had a positive effect on the supra-maximally LRH-stimulated secretion of LH and FSH as well as on the non-stimulated secretion of LH. EB had no effect on the non-stimulated secretion of FSH. After 5 days of in vivo pretreatment with LRH only, the in vitro non-stimulated and supra-maximally LRH-stimulated secretion of both LH and FSH were strongly impaired, the effect correlating well with the LRH-induced depletion of the pituitary LH/FSH stores. In such LRH-pretreated rats EB had on day 5 a negative effect on the (already depressed) LRH-stimulated secretion of LH (not on that of FSH). EB had no effect on the non-stimulated LH/FSH secretion. It could be demonstrated that the negative effect of the combined LRH/EB pretreatment was mainly due to the depressing effect of this treatment on the pituitary LH and FSH stores: the effect of oestradiol on the pituitary LRH-responsiveness (release as related to pituitary gonadotrophin content) remained positive. In LRH-pretreated rats, however, this positive effect of EB was smaller than in rats not pretreated with LRH. Four days after removal of the minipumps there was again a positive effect of EB on the LRH-stimulated secretion of LH and FSH as well as on the non-stimulated secretion of LH. The positive effect of EB on the pituitary LRH-responsiveness was as strong as in rats which had not been exposed to exogenous LRH. The non-stimulated secretion of FSH was again not affected by EB. The results demonstrate that the effect of EB on the oestrogen-sensitive components of gonadotrophin secretion consists of two components: an effect on the pituitary LRH-responsiveness proper, and an effect on the pituitary LH/FSH stores. The magnitude of the effect of EB on the LRH-responsiveness is LRH dependent: it is very weak (almost zero) in LRH-pretreated rats, but strong in rats not exposed to LRH as well as in rats of which the LRH-pretreatment was stopped 4 days previously. Similarly, the effect of EB on the pituitary LH and FSH stores is LRH-dependent: in the absence of LRH, EB has no influence on the contents of these stores, but EB can potentiate the depleting effect of LRH on the LH/FSH-stores. Also this effect disappear after cessation of the LRH-pretreatment.


Author(s):  
Т.А. Кuznetsova ◽  
N.А. Коlpakov

Показано, что длительное применение как минеральных, так и органических удобрений в различных дозах и сочетаниях положительно влияет на урожайность огурца. Установлено, что в условиях 14-й ротации урожайность на всех вариантах опыта была выше, чем на контрольном варианте и составила 16,4–22,3 т/га, против 16,0 т/га в контроле.The authors show that the long-term use of both mineral and organic fertilizers in different doses and combinations has a positive effect on the yield of cucumber. It was found that under 14 rotation conditions the yield on all variants of the experiment was higher than on the control variant and amounted to 16.4–22.3 t/ha, against 16.0 t/ha on the control.


GYNECOLOGY ◽  
2020 ◽  
Vol 22 (1) ◽  
pp. 14-18 ◽  
Author(s):  
Tatyana Yu. Pestrikova ◽  
Elena A. Yurasova ◽  
Igor V. Yurasov

Relevance. Endometriosis is a common gynecological disease that affects up to 10% of women of reproductive age worldwide and is the main cause of pain and infertility. Endometriosis is a disease, although it has been known for a long time, nevertheless, in many ways it represents terra incognita for modern medicine. Aim. Analysis of literature on the feasibility of long-term and the use of the drug dienogest 2 mg (Vizanne), which has a positive effect on the quality of life of patients with endometriosis. Materials and methods. To write this review, a search was made for domestic and foreign publications in Russian and international search engines (PubMed, eLibrary, etc.) over the past 13 years. The review included articles from peer-reviewed literature. Results. The review presents data on the difficulties of verifying the diagnosis of endometriosis due to a combination of this pathology with pain, infertility, abnormal uterine bleeding. The pathogenesis of the origin of endometriosis-associated pain is presented. The efficacy of the use of the drug dienogest (Vizanne), which has a powerful antiproliferative effect that reduces the main symptoms of endometriosis (pain, bleeding), is substantiated. The expediency of long-term and safe use of the drug dienogest (Vizanne), which has a positive effect on the quality of life of patients with endometriosis, has been proved. Conclusions. Numerous scientific publications confirm the feasibility of prolonged use of the drug dienogest (Vizanne), to achieve remission during endometriosis.


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