scholarly journals Biochemical changes in cultured foetal rat liver explants

1975 ◽  
Vol 150 (2) ◽  
pp. 269-273 ◽  
Author(s):  
P C MacDonnel ◽  
E Ryder ◽  
J A Delvalle ◽  
O Greengard

Liver explants from 20-day-old foetuses cultured for 48h in the absence of serum released 70% of their total soluble protein content into the medium. In the presence of serum this loss still amounted to 60%. The concentration of total particulate protein remained unchanged but there was some translocation of mitochondrial enzymes to the cytosol, and enzymes expected to increase during this stage of development failed to do so. The addition of cortisol plus glucagon (to serum-containing media) did not decrease the loss of total soluble protein from the explants but induced considerable tyrosine aminotransferase activity which was not released into the medium. The observations suggest that under the usual culture conditions a minority of the cells retain their functional integrity. The extent of deterioration, not reflected in histologically visible necrosis or cell damage, can be conveniently monitored by the malate dehydrogenase activity released to the medium.

2015 ◽  
Vol 52 (1-2) ◽  
pp. 19-36 ◽  
Author(s):  
Roya Razavizadeh

Abstract The effect of salicylic acid (SA) on the salt (NaCl) tolerance mechanism was studied in canola plants (oilseed rape, Brassica napus L.) by molecular and physiological experiments in plant tissue culture. Seeds of B. napus ‘Ocapy’ were germinated at 0, 50, and 100 mM NaCl on Murashige and Skoog (MS) medium containing different levels (0, 2, and 5 μM) of SA for 4 weeks. Total chlorophyll, carotenoid, and flavonoid content increased in response to interactive effects of SA and NaCl treatments at some concentrations. Proline content was increased under salt and SA treatments in shoot and root tissues. Salt alone and in combination with SA increased the total soluble protein content of shoots only, while the different concentrations of SA in the culture media affected variously the total soluble protein content. Protein patterns of shoots and roots showed some remarkable differences, based on gel electrophoresis and the consequent analysis of bands by ImageJ program. The relative expression of 15 and 12 protein bands in shoots and roots, respectively, differed under the applied treatments. In addition, the protein profile indicated that salinity and SA regulate the expression of salt-stress-inducible proteins as well as induced de novo synthesis of specific polypeptides. The findings may help to explain the salt tolerance mechanisms and to produce salt-tolerant canola plants.


2021 ◽  
Vol 17 (6) ◽  
pp. 805-817
Author(s):  
Sharifah Zafierah Syed Badrulzaman ◽  
Nurhana Nadia Ramlan ◽  
Munirah Adibah Kamarul Zaman ◽  
Azzreena Mohamad Azzeme

Musa paradisiaca cv Lang belongs to cooking banana group, and it has high potential to be used in banana chips production. Like other cultivars, M. paradisiaca cv Lang is susceptible towards water shortage, therefore affecting banana growth and productivity. In this study, to mimic the drought condition, pseudo-drought stress was given to in vitro Lang banana seedlings by adding polyethylene glycol (PEG). Overall, decrement of roots length and chlorophyll (Chl) content was displayed by the seedlings exposed to 1%, 2%, 3%, 4%, and 5% (w/v) of PEG after three weeks of exposure. The proline content, total soluble protein content, and antioxidant capacity in leaf and roots, however, countered differently towards different levels of drought. Proline content showed the highest in leaf of 2% (w/v) PEG-treated seedling (12.66±0.38 µmoles/g) while the total soluble protein content showed the highest in roots of 5% (w/v) of PEG-treated seedling (30.65±1.07 mg/g FW). Antioxidant capacity of stressed seedlings revealed the catalase (CAT), guaiacol peroxidase (POD), and ascorbate peroxidase (APX) activities were the highest in the leaf of 1% (w/v) (10.69±5.06 µmol/min/mg), 4% (w/v), (0.079±0.03 µmol/min/mg), and 5% (w/v) (9.11±8.47 µmol/min/mg) of PEG- treated seedlings, respectively. Meanwhile, the highest CAT, POD, and APX activities in the roots were determined in 3% (w/v) (0.49±0.04 µmol/min/mg), 2% (w/v) (0.03±0.02 µmol/min/mg), and 3% (w/v) (16.69±0.5 µmol/min/mg) of PEG-treated seedlings, respectively. These data show that PEG can be a priming agent to induce defense system at seedling stage of banana, which could enhance their survivability during ex vitro acclimatization.


Author(s):  
L. O. Sakhno ◽  
K. V. Lystvan

Aim. The study of possible unintended biochemical peculiarities of newly obtained herbicide-resistant canola (Brassica napus L.) plants expressing the lox-dependent BAR gene was the aim. Methods. Total soluble protein content, total free radical scavenging activity, and superoxide dismutase activity have been investigated using Bradford’s, DPPH, and nitroblue tetrazolium assays, respectively. Fresh weight of plants grown on media with or without phosphinothricin was also measured. Results. The antioxidant activity of leaf extracts of untransformed plants under in vitro growth condi-tions had no significant differences in comparison with ones of phospinothricin-resistant plants in the third generation. No significant changes in parameters investigated were observed in transgenic plants cultivated on media with herbicide addition compared to ones grown on media without it. Fresh weight and total soluble protein content were similar in transgenic and untransformed canola plants under growth without phosphinothricin. Conclusions. Lox-dependent BAR gene introduction and expression resulted in no significant differences in leaf antioxidant activity in transgenic canola plants comparing to untransformed controls.Keywords: Brassica napus, antioxidant activity, DPPH, glufosinate, superoxide dismutase.


2014 ◽  
Vol 42 (2) ◽  
pp. 446-452 ◽  
Author(s):  
Sevgi MARAKLI ◽  
Aslihan TEMEL ◽  
Nermin GOZUKIRMIZI

Potential alleviation effects of Homobrassinosteroid (HBR) (0.5 and 1 µM HBR) on root germination, cell division and antioxidant system enzymes (superoxide dismutase and catalase) of barley (Hordeum vulgare L. cv. ‘Hilal’) roots grown under different salt concentrations (150 mM and 250 mM) were investigated during 48 and 72 h at dark with their controls. Salt applications decreased primary root lengths, seminal root lengths, number of roots from one seed, mitotic activity and induced mitotic abnormalities. In addition, salt application decreased protein content but increased enzyme activities both at 48 h and 72 h when compared to control. Roots treated with HBR enhanced root lengths and root number. HBR-treated roots showed more mitotic activity, mitotic abnormalities and significant enlargements at the root tips when compared to controls and only salt-treated samples. Salt + HBR applications stimulated root lengths and showed more mitotic activity and mitotic abnormalities when compared to only salt-treated samples. Salt application increased superoxide dismutase and catalase activities both at 48 h and 72 h.  However, HBR application decreased total soluble protein content and increased enzyme activities especially at 48 h compared to control. Depending on concentration and timing, salt + HBR treatments showed varying results in total soluble protein content and enzyme activities. There was also a significant change in protein banding patterns of experimental groups separated by SDS-PAGE. These findings are expected to contribute to understanding of how HBR affect barley roots grown under different salt concentrations.


2014 ◽  
Vol 1061-1062 ◽  
pp. 166-169
Author(s):  
Na Gu ◽  
Jin Long Gao ◽  
Kui Tao Wang ◽  
Wen Cui Dong ◽  
Yu Lin Ma

ZnO/SnO2/montmorillonite that could flocculate and restrict the growth of Microcystis aeruginosa used as a probe of cyanobacterial was prepared and characterized by means of XRD and TEM. In ZnO/SnO2/montmorillonite, ZnO and SnO2nanoparticles disperse on montmorillonite surface. The determinations of chlorophyll a levels and total soluble protein content demonstrate that ZnO/SnO2/montmorillonite has stronger flocculation effect on Microcystis aeruginosa compared with natural montmorillonit under visible light, and has a good photocatalytic degradation effect on Microcystis aeruginosa under UV irradiation. Under UV, 96.79% removal efficiency is achieved for Microcystis aeruginosa in 1h using 50mg.L-1ZnO/SnO2/montmorillonite. The synergy of absorption flocculation and photocatalysis of ZnO/SnO2/montmorillonite promotes the removal of Microcystis aeruginosa.


1969 ◽  
Vol 115 (1) ◽  
pp. 19-24 ◽  
Author(s):  
Olga Greengard

1. The administration of glucagon, cAMP [adenosine 3′,5′-(cyclic)-monophosphate], BcAMP [6-N-2′-O-dibutyryladenosine 3′,5′-(cyclic)-monophosphate] or adrenaline to foetal rats during the last 2 days of gestation evoked the appearance of tyrosine aminotransferase and enhanced the accumulation of glucose 6-phosphatase in the liver. In foetuses 1–2 days younger only BcAMP was effective. After birth liver glucose 6-phosphatase no longer responds to glucagon or BcAMP. Tyrosine aminotransferase is still inducible by these agents in 2-day-old rats, but not in 50-day-old rats. After adrenalectomy of adults glucagon or BcAMP can enhance the induction of the enzyme by hydrocortisone. The results indicate that the ability to synthesize tyrosine aminotransferase and glucose 6-phosphatase when exposed to cAMP develops sooner than the ability to respond to glucagon with an increase in the concentration of cAMP; the responsiveness of enzymes to different hormones changes with age. A scheme illustrating the sequential development of competence in regulating the level of an enzyme is presented. 2. Actinomycin inhibited the effects of glucagon and BcAMP on liver tyrosine aminotransferase and glucose 6-phosphatase in foetal rats. Growth hormone, insulin and hydrocortisone did not enhance the formation of these enzymes. 3. The time-course of accumulation of glucose 6-phosphatase in the kidney is different from that in the liver. Hormones that increase the accumulation in foetal liver do not do so in the kidney of the same foetus or in the livers of postnatal rats.


2014 ◽  
Vol 9 (12) ◽  
pp. 1182-1195
Author(s):  
Samar Naseer ◽  
Tariq Mahmood

AbstractThe present study was designed to analyze genetically somaclonal variants using biochemical and molecular markers. Efficient tissue culture protocol for Solanum melongena L. cv. Nirrala was developed. Maximum callus induction (100%) was observed for Murashige and Skoog (MS) media supplemented with 2.0 mg L−1 naphthalene acetic acid +0.5 mg L−1 6-benzylaminopurine; and nodal explants gave best callusing response (88.8%) as compared to internodes (88.3%) and leaves (87.7%). The best shooting was induced on nodal and internodal callus in the presence of 2.0 mg L−1 6-benzylaminopurine. Total soluble protein content of callus and regenerated variant plants was estimated for biochemical analysis, and largest amount of soluble protein was found in callus (6.54 mg g−1 fresh tissue) followed by variant plant grown on 2.0 mg L−1 6-benzylaminopurine (5.96 mg g−1 fresh tissue). Random amplification of polymorphic DNA technique was done with five decamer primers (OPC1-OPC5) and maximum polymorphism was detected by OPC 2 (26.99%) among all samples, whereas nodal callus on media containing 1.0 mg L−1 naphthalene acetic acid +1.0 mg L−1 6-benzylaminopurine showed highest polymorphism producing 22 bands, out of which 8 bands were polymorphic. The study shows that this marker system can provide better evaluation of genetic variation induced by tissue culture.


2000 ◽  
Vol 8 (3) ◽  
pp. 241-287 ◽  
Author(s):  
GM Jones

The transfer of a blastocyst established the first human clinical pregnancy following in vitro fertilization (IVF). Nine years later Cohen et al. reported pregnancies resulting from the transfer of cryopreserved human blastocysts. However, it was another six years before the first report of births resulting from the transfer of human blastocysts produced in vitro appeared in the medical literature. In the intervening period clinics have opted to transfer embryos at the early cleavage stage to the uterus, despite the fact that in vivo the embryo does not enter the uterus until two to three days later at the morula to blastocyst stage of development. The viability and potential for implantation of blastocysts is high, as indicated by the finding that more than 60% of in-vivo-derived blastocysts, recovered by uterine lavage following artificial insemination of fertile donors, implant and develop into viable fetuses when transferred to recipients. This is in stark contrast to the 10–20% of in-vitro-produced embryos transferred at the early cleavage stage of development that result in a live-birth. This reduction in viability following transfer of in-vitro-derived early cleavage stage embryos may have several possible explanations: (1) a failure of implantation due to poor synchronization between the embryo and the uterine endometrium; (2) a hostile environment in the uterus for early cleavage stage embryos; (3) sub-optimal in vitro culture conditions which result in a reduction in embryo viability; (4) the assumption that all oocytes retrieved in an IVF cycle have an equal ability to develop into viable embryos; and (5) the failure to identify the most viable embryo in a cohort. Certainly, improving culture conditions and laboratory techniques for developing high quality blastocysts routinely in vitro will not only address many of the above questions but will also improve the quality and viability of earlier stages of embryo development.


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