Effect of polyunsaturated fatty acids on diphenyl hydantoin-induced genetic damage in vitro and in vivo

Phenytoin sodium/diphenyl hydantoin (DPH) is used by a major segment of epileptics and neuro surgery patients with head injury to prevent seizures. DPH is a known mutagen, carcinogen, and teratogen. Essential fatty acids (EFAs) are critical for various tissues and were reported to act as anti-mutagenic agents. In the present study we assessed the effect of five EFAs on DPH-induced genetic damage both in vitro and in vivo. DPH induced significant genetic damage. Of all the EFAs (linoleic acid, α-linolenic acid, gamma-linolenic acid, arachidonic acid, dihomo-gamma-linolenic acid, and eicosapentaenoic acid) studied, all except eicosapentaenoic acid showed significant decrease in DPH induced genetic damage as assessed by micronucleus (MN) test. However, gamma-linolenic acid (GLA) was found to be the most effective in reducing the number of MN containing lymphocytes both in vitro and in vivo to control values. EFAs when tested alone produced insignificant increase in the amount of genetic damage but when tested in combination with DPH the number of micronuclei containing lymphocytes was reduced; but the DNA ladder pattern, an indication of DNA damage, was increased. This apparently paradoxical action of EFAs, especially of GLA, suggests that, in all probability, fatty acids induce apoptosis of cells that harbor significant DNA damage. Based on these results we suggest that GLA functions as a unique endogenous molecule that protects cells from accumulating genetic damage.     

Changes in growth and photosynthetic characteristics of Ocimum sanctum under growth regulator treatments

A field experiment was conducted to investigate the effect of growth regulators on growth characteristics such as root length, shoot length, total leaf area, number of inflorescence per plant, number of flower per inflorescence, whole plant fresh weight and whole plant dry weight. Photosynthetic characteristics were also analyzed based on the same experiment. For this, various photosynthetic pigment contents such as chlorophyll, carotenoid, anthocyanin and xanthophyll content were calculated. The conventional growth regulator abscisic acid (ABA) and non-conventional growth regulator triazole compound paclobutrazol (PBZ) were used. Root length increased due to growth regulator treatment, but shoot length decreased. Leaf area was decreased due to growth regulator treatment. The number of inflorescence increased in ABA treated plants, but it was decreased in PBZ treated plants. In ABA treated plants, the number of flowers per inflorescence was increased. In PBZ treated plants the number of inflorescence was reduced. The whole plant fresh weight (FW) and dry weight (DW) were increased in ABA and PBZ treated plants. There was an increase in chlorophyll content in growth regulator treated plants compared to control, and it was more in PBZ treated plants. The carotenoid content was also increased in ABA and PBZ treated plants.     

Alterations in sucrose metabolizing enzyme activities and total phenol content of Curcuma longa L. as affected by different triazole compounds

Changes in the sucrose metabolism of Curcuma longa L. plants were studied under treatment with different triazole compounds viz., triadimefon (TDM) and propiconazole (PCZ). Plants were treated with TDM at 15 mg/L and PCZ at 10 mg/L separately by soil drenching on 80, 110, and 140 days after planting (DAP). The plants were harvested randomly on 90, 120, and 150 DAP to determine the effect of both the triazoles on sucrose metabolizing enzymes and phenol content. The sucrose metabolism was studied by analyzing sucrose metabolizing enzymes like sucrose synthase and sucrose phosphate synthase. All the analyses were assayed in leaves and tubers of both control and treated plants. It was found that both of the triazole compounds had profound effects on these parameters.     

ABA and GA3 affect the growth and pigment composition in Andrographis paniculata Wall.ex Nees., an important folk herb

In this study, 5 μmol·L⁻¹ abscisic acid (ABA) and gibberellic acid (GA₃) were used to study the effect of both growth regulators on the morphological parameters and pigment composition of Andrographis paniculata. The growth regulators were applied by means of foliar spray during morning hours. ABA treatment inhibited the growth of the stem and internodal length when compared with control, whereas GA₃ treatment increased the plant height and internodal length. The total number of leaves per plant decreased in the ABA-treated plants, but GA₃ treatment increased the total number of leaves when compared with the control. Both growth regulators (ABA and GA₃) showed increased leaf area. ABA and GA₃ treatments slightly decreased the total root growth at all the stages of growth. The growth regulator treatments increased the whole plant fresh and dry weight at all stages of growth. ABA enhanced the fresh and dry weight to a larger extent when compared with GA₃. An increase in the total chlorophyll content was recorded in ABA and GA₃ treatments. The chlorophyll-a, chlorophyll-b, and carotenoids were increased by ABA and GA₃ treatments when compared with the control plants. The xanthophylls and anthocyanin content were increased with ABA and GA₃ treatments in A. paniculata plants.     

Alterations in sucrose metabolizing enzyme activities and total phenol content of Curcuma longa L. as affected by different triazole compounds

Changes in the sucrose metabolism of Curcuma longa L. plants were studied under treatment with different triazole compounds viz., triadimefon (TDM) and propiconazole (PCZ). Plants were treated with TDM at 15 mg/L and PCZ at 10 mg/L separately by soil drenching on 80, 110, and 140 days after planting (DAP). The plants were harvested randomly on 90, 120, and 150 DAP to determine the effect of both the triazoles on sucrose metabolizing enzymes and phenol content. The sucrose metabolism was studied by analyzing sucrose metabolizing enzymes like sucrose synthase and sucrose phosphate synthase. All the analyses were assayed in leaves and tubers of both control and treated plants. It was found that both of the triazole compounds had profound effects on these parameters.     

Changes in growth and photosynthetic characteristics of Ocimum sanctum under growth regulator treatments

A field experiment was conducted to investigate the effect of growth regulators on growth characteristics such as root length,shoot length,total leaf area,number of inflorescence per plant,number of flower per inflorescence,whole plant fresh weight and whole plant dry weight.Photosynthetic characteristics were also analyzed based on the same experiment.For this,various photosynthetic pigment contents such as chlorophyll,carotenoid,anthocyanin and xanthophyll content were calculated.The conventional growth regulator abscisic acid (ABA) and non-conventional growth regulator triazole compound paclobutrazol (PBZ) were used.Root length increased due to growth regulator treatment,but shoot length decreased.Leaf area was decreased due to growth regulator treatment.The number of inflorescence increased in ABA treated plants,but it was decreased in PBZ treated plants.In ABA treated plants,the number of flowers per inflorescence was increased.In PBZ treated plants the number of inflorescence was reduced.The whole plant fresh weight (FW) and dry weight (DW) were increased in ABA and PBZ treated plants.There was an increase in chlorophyll content in growth regulator treated plants compared to control,and it was more in PBZ treated plants.The carotenoid content was also increased in ABA and PBZ treated plants.     

A constitutively expressed 36 kDa exochitinase from Bacillus thuringiensis HD-1

A 36 kDa chitinase was purified by ion exchange and gel filtration chromatography from the culture supernatant of Bacillus thuringiensis HD-1. The chitinase production was independent of the presence of chitin in the growth medium and was produced even in the presence of glucose. The purified chitinase was active at acidic pH, had an optimal activity at pH 6.5, and showed maximum activity at 65 degrees C. Of the various substrates, the enzyme catalyzed the hydrolysis of the disaccharide 4-MU(GlnAc)(2) most efficiently and was therefore classified as an exochitinase. The sequence of the tryptic peptides showed extensive homology with Bacillus cereus 36 kDa exochitinase. The 1083 bp open reading frame encoding 36 kDa chitinase was amplified with primers based on the gene sequence of B. cereus 36 kDa exochitinase. The deduced amino-acid sequence showed that the protein contained an N-terminal signal peptide and consisted of a single catalytic domain. The two conserved signature sequences characteristic of family 18 chitinases were mapped at positions 105-109 and 138-145 of Chi36. The recombinant chitinase was expressed in a catalytically active form in Escherichia coli in the vector pQE-32. The expressed 36 kDa chitinase potentiated the insecticidal effect of the vegetative insecticidal protein (Vip) when used against neonate larvae of Spodoptera litura.     

Identification of the sucrose non-fermenting related kinase SNRK, as a novel LKB1 substrate

Recent work has shown that the LKB1 tumour suppressor protein kinase phosphorylates and activates protein kinases belonging to the AMP activated kinase (AMPK) subfamily. In this study, we identify the sucrose non-fermenting protein (SNF1)-related kinase (SNRK), a largely unstudied AMPK subfamily member, as a novel substrate for LKB1. We demonstrate that LKB1 activates SNRK by phosphorylating the T-loop residue (Thr173), and that the LKB1 regulatory subunits STRAD and MO25 are required for LKB1 to activate SNRK. We find that SNRK is not active when expressed in HeLa cells that lack expression of LKB1, and its activity is restored by expression of wild type LKB1, but not catalytically deficient LKB1. We also present evidence that two other AMPK-related kinases more distantly related to AMPK than SNRK, namely NIM1 and testis-specific serine/threonine kinase-1 (TSSK1) are not substrates for LKB1. Tissue distribution analysis indicates that SNRK protein is mainly expressed in testis, similar to TSSK isoforms, whereas NIM1 is more widely expressed. These results provide evidence that SNRK could mediate some of the physiological effects of LKB1.     

Cultural stability ofStreptomyces fradiae in the production of xylose isomerase: studies in shake flasks

Comprehensive studies of pure colonies of Streptomyces fradiae in the production of xylose isomerase by submerged fermentation at shake flask level revealed poor culture stability with respect to enzyme production, biomass formation, degree of pigmentation, quantity of glucose and xylose utilization, level of enzyme in cell-free culture broth and final pH of the fermentation medium. The results serve to stress obligatory evaluation of culture stability of Streptomyces strains in determining their suitability for use in developing fermentation processes for commercial exploitation.