scholarly journals Thioredoxin peroxidases can foster cytoprotection or cell death in response to different stressors: over- and under-expression of thioredoxin peroxidase in Drosophila cells

2003 ◽  
Vol 371 (3) ◽  
pp. 743-752 ◽  
Author(s):  
Svetlana N. RADYUK ◽  
Rajindar S. SOHAL ◽  
William C. ORR
Keyword(s):  
Hydrogen Peroxide ◽  
Heat Stress ◽  
S2 Cells ◽  
Free Radical Biol ◽  
Drosophila Cell ◽  
Functional Roles ◽  
Thioredoxin Peroxidase ◽  
Negative Effect ◽  
Drosophila Cells

Recently, we identified a set of five genes constituting the peroxiredoxin gene family in Drosophila melanogaster [Radyuk, Klichko, Spinola, Sohal and Orr (2001) Free Radical Biol. Med. 31, 1090–1100]. This set includes two abundant thioredoxin peroxidase (TPx) species, namely Drosophila peroxiredoxin DPx-4783, a cytosolic TPx and DPx-5037, a mitochondrial TPx. Overexpression of either one of them in Drosophila S2 cells conferred increased resistance to toxicity induced by hydrogen peroxide, paraquat or cadmium. To understand further the functional roles of these enzymes in vivo, we report in the present study the effects of decreased expression, using RNA interference, on the response of S2 cells to different stressors. When either of the TPxs was blocked, cells became relatively more susceptible to oxidative stress caused by exposure to hydrogen peroxide or paraquat, but were unaffected when challenged with copper and heat stress. In contrast, TPx overexpressing cells were more susceptible to copper and heat stress when compared with control cells and exhibited DNA fragmentation. Furthermore, when cells were supplemented with N-acetyl-l-cysteine together with copper, there was a clear negative effect on cell survival, which was exacerbated by TPx overexpression. Manipulations in the levels of TPxs demonstrated that, under different stress conditions, these enzymes might have both beneficial and detrimental effects on Drosophila cell viability.

scholarly journals A Multicomponent Assembly Pathway Contributes to the Formation of Acentrosomal Microtubule Arrays in Interphase Drosophila Cells

2008 ◽  
Vol 19 (7) ◽  
pp. 3163-3178 ◽  
Author(s):  
Gregory C. Rogers ◽  
Nasser M. Rusan ◽  
Mark Peifer ◽  
Stephen L. Rogers
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Cell Types ◽  
Microtubule Assembly ◽  
Animal Cells ◽  
Drosophila Cell ◽  
Assembly Pathway ◽  
Interphase Cells ◽  
Drosophila Cells

In animal cells, centrosomes nucleate microtubules that form polarized arrays to organize the cytoplasm. Drosophila presents an interesting paradox however, as centrosome-deficient mutant animals develop into viable adults. To understand this discrepancy, we analyzed behaviors of centrosomes and microtubules in Drosophila cells, in culture and in vivo, using a combination of live-cell imaging, electron microscopy, and RNAi. The canonical model of the cycle of centrosome function in animal cells states that centrosomes act as microtubule-organizing centers throughout the cell cycle. Unexpectedly, we found that many Drosophila cell-types display an altered cycle, in which functional centrosomes are only present during cell division. On mitotic exit, centrosomes disassemble producing interphase cells containing centrioles that lack microtubule-nucleating activity. Furthermore, steady-state interphase microtubule levels are not changed by codepleting both γ-tubulins. However, γ-tubulin RNAi delays microtubule regrowth after depolymerization, suggesting that it may function partially redundantly with another pathway. Therefore, we examined additional microtubule nucleating factors and found that Mini-spindles, CLIP-190, EB1, or dynein RNAi also delayed microtubule regrowth; surprisingly, this was not further prolonged when we codepleted γ-tubulins. Taken together, these results modify our view of the cycle of centrosome function and reveal a multi-component acentrosomal microtubule assembly pathway to establish interphase microtubule arrays in Drosophila.

scholarly journals The impact of oxidative stress on the neurotoxic effect of acetaminophen

2019 ◽  
Vol 59 (9) ◽  
pp. 106-109
Author(s):  
Ksenia A. Zagorodnikova ◽  
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Neurotoxic Effect ◽  
Mtt Method ◽  
Culture Viability ◽  
Negative Effect ◽  
High Concentrations ◽  
Pharmacologically Active ◽  
The Impact

Paracetamol (acetaminophen, APAP) is a commonly-used antipyretic and analgesic. However, there have been reports indicating possible link between its use in pregnancy and impaired neuropsychic development in children. A number of prospective studies of the possible negative effect of acetaminophen on the development of a child after his mother took this drug during pregnancy, as well as the results of studies on glioma cells and neurons in murine cortex, may indicate presence of the neurotoxic effect of acetaminophen. It is currently unclear if paracetamol itself being pharmacologically active neurotropic substance, or its metabolites, one of which – NAPQI (N-acetyl-p-benzoquinone imine) known by its toxic effects in mitochindria, play the most significant role in proposed neurotoxicity. Therefore it seems important to study each metabolite separately. The ability of acetaminophen(paracetamol) in concentrations of 1 mg/ml and 2 mg/ml to reduce cell viability was shown on cells of the PC12 neuronal line using MTT-method, which is based on the ability of mitochondria of viable cells to restore formazan 3-(4,5-dimethylthiazole)-2,5-diphenyl-2-tetrazolium bromide (MTT). Concentrations of 0.125 mg/ml, 0.25 mg/ml and 0.5 mg/ml had no similar impact on cell culture viability. In addition, the impact of hydrogen peroxide (as an inducer of oxidative stress) on the neurotoxic effect of acetaminophen was studied. We demonstrated that in the presence of 0.3 mM or 0.5 mM hydrogen peroxide and acetaminophen in concentrations of 1 mg/ml and 2 mg/ml reliably reduced the percentage of surviving cells. We showed that the decrease of the viability of the cells of the PC12 neuronal line is obvious only after exposure to high concentrations of acetaminophen, especially in the presence of hydrogen peroxide, which means that neurotoxic effect is not likely to occur in vivo.

scholarly journals Genetic control of kinetochore-driven microtubule growth in Drosophila mitosis

2021 ◽  
Author(s):  
Julia V. Popova ◽  
Gera A. Pavlova ◽  
Alyona V. Razuvaeva ◽  
Lyubov A. Yarinich ◽  
Evgeniya N. Andreyeva ◽  
...  
Keyword(s):  
Reverse Genetics ◽  
S2 Cells ◽  
Drosophila S2 Cells ◽  
Conserved Genes ◽  
Negative Effect ◽  
Microtubule Growth ◽  
Mitotic Spindle Assembly ◽  
Bipolar Spindles ◽  
Drosophila Cells ◽  
Dependent Pathway

Centrosome-containing cells assemble their spindles exploiting three main classes of microtubules (MTs): MTs nucleated by the centrosomes, MTs generated near the chromosomes/kinetochores, and MTs nucleated within the spindle by the augmin-dependent pathway. Mammalian and Drosophila cells lacking the centrosomes generate MTs at kinetochores and eventually form functional bipolar spindles. However, the mechanisms underlying kinetochore-driven MT formation are poorly understood. One of the ways to elucidate these mechanisms is the analysis of spindle reassembly following MT depolymerization. Here, we used an RNA interference (RNAi)-based reverse genetics approach to dissect the process of kinetochore-driven MT regrowth (KDMTR) after colcemid-induced MT depolymerization. This MT depolymerization procedure allows a clear assessment of KDMTR, as colcemid disrupts centrosome-driven MT regrowth but allows KDMTR. We examined KDMTR in normal Drosophila S2 cells and in S2 cells subjected to RNAi against conserved genes involved in mitotic spindle assembly: mast/orbit/chb (CLASP1), mei-38 (TPX2), mars (HURP), dgt6 (HAUS6), Eb1 (MAPRE1/EB1), Patronin (CAMSAP2), asp (ASPM) and Klp10A (KIF2A). RNAi-mediated depletion of Mast/Orbit, Mei-38, Mars, Dgt6 and Eb1 caused a significant delay in KDMTR, while loss of Patronin had a milder negative effect on this process. In contrast, Asp or Klp10A deficiency increased the rate of KDMTR. These results coupled with the analysis of GFP-tagged proteins (Mast/Orbit, Mei-38, Mars, Eb1, Patronin and Asp) localization during KDMTR suggested a model for kinetochore-dependent spindle reassembly. We propose that kinetochores capture the plus ends of MTs nucleated in their vicinity and that these MTs elongate at kinetochores through the action of Mast/Orbit. The Asp protein binds the MT minus ends since the beginning of KDMTR, preventing excessive and disorganized MT regrowth. Mei-38, Mars, Dgt6, Eb1 and Patronin positively regulate polymerization, bundling and stabilization of regrowing MTs until a bipolar spindle is reformed.

Fluorescent proteins for in vivo imaging, where's the biliverdin?

2020 ◽  
Vol 48 (6) ◽  
pp. 2657-2667
Author(s):  
Felipe Montecinos-Franjola ◽  
John Y. Lin ◽  
Erik A. Rodriguez
Keyword(s):  
Hydrogen Peroxide ◽  
In Vivo Imaging ◽  
Near Infrared ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Fluorescent Imaging ◽  
Infrared Light ◽  
Red Fluorescent Protein ◽  
Color Palette

Noninvasive fluorescent imaging requires far-red and near-infrared fluorescent proteins for deeper imaging. Near-infrared light penetrates biological tissue with blood vessels due to low absorbance, scattering, and reflection of light and has a greater signal-to-noise due to less autofluorescence. Far-red and near-infrared fluorescent proteins absorb light >600 nm to expand the color palette for imaging multiple biosensors and noninvasive in vivo imaging. The ideal fluorescent proteins are bright, photobleach minimally, express well in the desired cells, do not oligomerize, and generate or incorporate exogenous fluorophores efficiently. Coral-derived red fluorescent proteins require oxygen for fluorophore formation and release two hydrogen peroxide molecules. New fluorescent proteins based on phytochrome and phycobiliproteins use biliverdin IXα as fluorophores, do not require oxygen for maturation to image anaerobic organisms and tumor core, and do not generate hydrogen peroxide. The small Ultra-Red Fluorescent Protein (smURFP) was evolved from a cyanobacterial phycobiliprotein to covalently attach biliverdin as an exogenous fluorophore. The small Ultra-Red Fluorescent Protein is biophysically as bright as the enhanced green fluorescent protein, is exceptionally photostable, used for biosensor development, and visible in living mice. Novel applications of smURFP include in vitro protein diagnostics with attomolar (10−18 M) sensitivity, encapsulation in viral particles, and fluorescent protein nanoparticles. However, the availability of biliverdin limits the fluorescence of biliverdin-attaching fluorescent proteins; hence, extra biliverdin is needed to enhance brightness. New methods for improved biliverdin bioavailability are necessary to develop improved bright far-red and near-infrared fluorescent proteins for noninvasive imaging in vivo.

The depressing (negative) effect of oestradiol benzoate on the in vitro secretion of LH and FSH by the pituitary gland of the LRH-pretreated long-term ovariectomized rat changes into the augmentative (positive) effect after discontinuation of the LRH-pretreatment

1985 ◽  
Vol 110 (3) ◽  
pp. 329-337 ◽  
Author(s):  
G. A. Schuiling ◽  
H. Moes ◽  
T. R. Koiter
Keyword(s):  
Depressing Effect ◽  
Ovx Rats ◽  
Gonadotrophin Secretion ◽  
Oestradiol Benzoate ◽  
Negative Effect ◽  
Positive Effect ◽  
Perifusion System

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.

Pengaruh Luka Iris Pada Tikus Dengan Paparan Hidrogen Peroxida 3% terhadap Epitelisasi dan Pembentukan Eksudat pada Permukaan Epitel Kulit.

2020 ◽  
Vol 17 (2) ◽  
pp. 172
Author(s):  
HARMAN AGUSAPUTRA ◽  
MARIA SUGENG ◽  
AYLY SOEKAMTO ◽  
ATIK WULANDARI
Keyword(s):  
Hydrogen Peroxide ◽  
Wound Healing ◽  
The Other ◽  
Delayed Wound Healing ◽  
Peroxide Group ◽  
Hemostatic Effect ◽  
Significant Difference ◽  
Negative Effect ◽  
Wound Healing Effect ◽  
Epithelial Growth

<p><strong>Abstract</strong></p><p><strong>Background:</strong> Hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) as antiseptic has been used frequently to clean woundsin in hospitals and clinics. Hydrogen peroxide has the effectof strong oxidative that can kill pathogens. It can clean up debris and necrotic tissuesin wounds. Hydrogen peroxidealso has hemostatic effect that can help to stop bleeding. Besides antiseptic effects, hydrogen peroxide i s suspected of having negative effect in wound healing. Hydrogen peroxide presumably could cause delayed wound healing by exudate formation and delayed epithelial growth.</p><p><strong>Method</strong>: This study was conducted in the laboratory using 48 white mice that were divided into 2 groups. All the mice were purposely wounded. Afterwards in one group the wounds were clean up using hydrogen peroxide, while in the other group without hydrogen peroxide as control. The wounds of both groups were observed on day 1, day 3 and day 7. On day 1 and day 3, both groups did not show significant difference.</p><p><strong>R</strong><strong>esult</strong> : on day 7 showed that the wound healing in hydrogen peroxide group were delayed. Fifty percent of them had the formation of exudate and 62.5% of them showed delayed epithelial growth.</p><p><strong>Conclusion </strong>: This study could show hydrogen peroxide as wound antiseptic has delayed wound healing effect.</p><p><strong>Keyword</strong>: hydrogen peroxide, wound healing</p>

Associated microflora of medicinal ferns: biotechnological potentials and possible applications

2016 ◽  
Vol 5 (03) ◽  
pp. 4927 ◽  
Author(s):  
Shubhi Srivastava ◽  
Paul A. K.
Keyword(s):  
Secondary Metabolites ◽  
Growth Promotion ◽  
Biological Activities ◽  
Hydrolytic Enzymes ◽  
In Vitro Production ◽  
Wide Range ◽  
Negative Effect ◽  
Bacteria And Fungi

Plant associated microorganisms that colonize the upper and internal tissues of roots, stems, leaves and flowers of healthy plants without causing any visible harmful or negative effect on their host. Diversity of microbes have been extensively studied in a wide variety of vascular plants and shown to promote plant establishment, growth and development and impart resistance against pathogenic infections. Ferns and their associated microbes have also attracted the attention of the scientific communities as sources of novel bioactive secondary metabolites. The ferns and fern alleles, which are well adapted to diverse environmental conditions, produce various secondary metabolites such as flavonoids, steroids, alkaloids, phenols, triterpenoid compounds, variety of amino acids and fatty acids along with some unique metabolites as adaptive features and are traditionally used for human health and medicine. In this review attention has been focused to prepare a comprehensive account of ethnomedicinal properties of some common ferns and fern alleles. Association of bacteria and fungi in the rhizosphere, phyllosphere and endosphere of these medicinally important ferns and their interaction with the host plant has been emphasized keeping in view their possible biotechnological potentials and applications. The processes of host-microbe interaction leading to establishment and colonization of endophytes are less-well characterized in comparison to rhizospheric and phyllospheric microflora. However, the endophytes are possessing same characteristics as rhizospheric and phyllospheric to stimulate the in vivo synthesis as well as in vitro production of secondary metabolites with a wide range of biological activities such as plant growth promotion by production of phytohormones, siderophores, fixation of nitrogen, and phosphate solubilization. Synthesis of pharmaceutically important products such as anticancer compounds, antioxidants, antimicrobials, antiviral substances and hydrolytic enzymes could be some of the promising areas of research and commercial exploitation.

scholarly journals A Dominant Negative Effect of eth-1r, a Mutant Allele of the Neurospora crassa S-Adenosylmethionine Synthetase-Encoding Gene Conferring Resistance to the Methionine Toxic Analogue Ethionine

Genetics ◽  
1996 ◽  
Vol 144 (4) ◽  
pp. 1455-1462
Author(s):  
José L Barra ◽  
Mario R Mautino ◽  
Alberto L Rosa
Keyword(s):  
Neurospora Crassa ◽  
Dominant Negative ◽  
Dominant Negative Effect ◽  
Resistant Phenotype ◽  
Negative Effect ◽  
Encoding Gene ◽  
Adenosylmethionine Synthetase ◽  
Adomet Synthetase ◽  
Identical Gene

eth-1r a thermosensitive allele of the Neurospora crassa S-adenosylmethionine (AdoMet) synthetase gene that confers ethionine resistance, has been cloned and sequenced. Replacement of an aspartic amino acid residue (D48 → N48), perfectly conserved in prokaryotic, fungal and higher eukaryotic AdoMet synthetases, was found responsible for both thermosensitivity and ethionine resistance conferred by eth-1r. Gene fusion constructs, designed to overexpress eth-1r in vivo, render transformant cells resistant to ethionine. Dominance of ethionine resistance was further demonstrated in eth-1  +/eth-1r partial diploids carrying identical gene doses of both alleles. Heterozygous eth-1  +/eth-1r cells have, at the same time, both the thermotolerance conferred by eth-1  + and the ethionine-resistant phenotype conferred by eth-1r. AdoMet levels and AdoMet synthetase activities were dramatically decreased in heterozygous eth-1  +/eth-1r cells. We propose that this negative effect exerted by eth-1r results from the in vivo formation of heteromeric eth-1  +/eth-1r AdoMet synthetase molecules.

An updated review on behavior of domestic quail with reference to the negative effect of heat stress

2021 ◽  
pp. 1-14
Author(s):  
Fiza Batool ◽  
Rana M. Bilal ◽  
Faiz Ul Hassan ◽  
Taquir A. Nasir ◽  
Majid Rafeeque ◽  
...  
Keyword(s):  
Heat Stress ◽  
Negative Effect
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