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Abstract

One of the negative aspects of the intensive use of herbicides is related to the selection of resistant biotypes (Gasquez 2001). Of all biotypes resistant to herbicides, 93 species do not respond to sulfonylurea herbicides (ALS-inhibiting herbicides). The acetolactate synthase (ALS) enzyme is the first step in biosynthesis of a branched chain amino acids (valine, leucine, isoleucine). In Poland the problem of resistance to sulfonylurea herbicides has been discussed since 2001 (Rola and Marczewska 2002). Resistance tests of Apera spica-venti biotypes were conducted in the greenhouse conditions. Chlorsulfuron was applied at the four-leaf stage of development at rates ranging from 11.25 to 360 g/ha. In confirmation of resistance to chlorsulfuron as identified in biological tests, the chemical analyses were performed. The analyses investigated the influence of different doses of chlorsulfuron on free amino acids content in the aboveground part of resistant and susceptible Apera spica-venti. The analyses were carried out applying high performance liquid chromatography method (HPLC). The resistance of the biotype was confirmed in amino acids analysis. In the resistant biotype followed the increase of valine, leucine and isoleucine concentration in comparison with untreated plants and those susceptible to chlorsulfuron biotype.

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Authors and Affiliations

Katarzyna Marczewska
Jerzy Sadowski
Henryka Rola
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Abstract

White adipose tissue plays an important role in the catabolism of branched chain amino acids (BCAAs). Two initial regulatory steps in BCAAs catabolism are catalyzed by branched chain aminotrans-ferase (BCAT) and branched chain α-keto acid dehydrogenase complex (BCKDH complex), respectively. It has been demonstrated that synthetic ligands for PPARγ receptors increased mRNA levels for enzymes involved in BCAAs catabolism. We hypothesized that feeding rats with diet rich in linoleic acid (LA), a natural PPARγ agonist modifies mRNA levels for enzymes catalyzing BCAAs degradation in adipose tissue. The current pilot study was aimed at the investigation of the effect of diet rich in LA on mRNA levels for BCATm, branched chain α-keto acid dehydrogenase (E1 component of the BCKDH), and mRNA levels for the regulatory enzymes of BCKDH complex, a specific kinase (BDK) and a specific phosphatase (PPM1K) in epididymal white adipose tissue (eWAT). Wistar male rats were fed with high unsaturated fat diet containing mainly linoleic acid (study group) or with the high saturated fat diet (control group). The relative mRNA levels were quantified by reverse transcription PCR. We have found that in rats fed diet rich in LA mRNA level for BCATm decreased, while mRNA amount for BDK increased. There was no difference between mRNA levels for BCKDH E1 and PPM1K. It is con-ceivable that changes in mRNA levels for enzymes involved in BCAAs metabolism in eWAT may lead to modification of BCAAs catabolic rate. Further studies are required to fully elucidate this issue.
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Authors and Affiliations

Małgorzata Knapik-Czajka
1
Justyna Bieleń
1
Monika Zajonz
1
Anna Gawędzka
1
Jagoda Drąg
1
Małgorzata Belczyk
1

  1. Department of Biochemical Analytics, Jagiellonian University Medical College, Kraków, Poland
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Abstract

Undisturbed branched-chain amino acids (BCAA) catabolism is necessary for normal heart function. The key enzyme in BCAA catabolism is a multienzyme branched-chain α-keto acid dehydro-genase complex (BCKDH). BCKDH activity is regulated mainly by reversible dephosphorylation (activa-tion)/phosphorylation (inactivation) cycle catalyzed by regulatory enzymes, a specific phosphatase (PPM1K) and kinase (BDK). 5-fluorouracil (5-FU) is widely used in the treatment of different types of cancer. 5-FU has the potential to cause a wide spectrum of cardiotoxicity, ranging from asymptomatic electrocardiographic changes to cardiomyopathy and subsequent cardiac failure. We hypothesize that 5-FU modifies BCKDH activity and affects cardiac muscle metabolism. The current study was aimed at the investigation of the in vivo effect of 5-FU on BCKDH activity and mRNA levels for E1, PPM1K and BDK. Wistar male rats were administered with 4 doses of 5-FU, 150 mg/kg b.wt. each (study group) or 0.3% methylcellulose (control group). BCKDH activity was assayed spectrophotometrically. The mRNA levels were quantified by real-time PCR. 5-FU treatment caused an increase in BCKDH activity that appears to result mainly from increased dephosphorylation of the complex and is associated with an increase of PPM1K mRNA level and reduction of BDK and E1 mRNA levels. It is conceivable that 5-FU stimulation of BCKDH is an adaptive reaction with the purpose of enhancing the BCAA catabolism and protecting from toxic effect caused by excessive accumulation of these amino acids in heart.
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Authors and Affiliations

Małgorzata Knapik-Czajka
1
Michał Jurczyk
2
Justyna Bieleń
1
Veronika Aleksandrovych
2
Anna Gawędzka
1
Paulina Stach
2
Jagoda Drąg
1
Krzysztof Gil
3

  1. Department of Biochemical Analytics, Jagiellonian University Medical College, Kraków, Poland
  2. Department of Pathophysiology, Jagiellonian University Medical College, Kraków, Poland
  3. 2Department of Pathophysiology, Jagiellonian University Medical College, Kraków, Poland

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