The gene expression of amylase, trypsin, and lipase in the digestive organs of the two-spotted cricket (Gryllus bimaculatus) was tested to understand how it overcomes starvation. Amylase gene expression in the foregut was reduced by digesting no food until starvation-3 days. Although that expression persisted to starvation-6 days, it returned to normal at refeeding-2 days. The expression of trypsin peaked at around 8 times as starvation started and at around 4 times at starvation-3 days. After refeeding, trypsin expression rose up to 14 times and then fell back to normal as feeding continued. Lipase gene expression remained elevated at 1.5-2 times when starvation started and returned to normal at refeeding-2 days. In the midgut, amylase expression decreased until starvation-3 days, increasing to about 2 times at starvation-6 days; it did not rise again by refeeding. Trypsin was constantly expressed regardless of starvation and refeeding, while lipase expression was reduced by 0.6-0.7 times by starvation and refeeding. Amylase gene expression in the hindgut was 0.2-0.3 times lower than starvation-6 days, and it increased by 0.5 times on refeeding-1 day and more than 1.5 times on refeeding-3 days. The gene expression of trypsin was almost identical to amylase.
This study was performed to discriminate defective loci by detection of congenital genetic disorder, to offer basic data for selection and improvement of Korean dairy cattle using frozen semen of Holstein bulls(16 proven and 93 candidate). The results obtained were as follows ; By the detection of DUMP(deficiency of uridine monophophate synthase) for 109 Holstein bulls(16 proven and 93 candidate), DUMP carrier was not found in whole animals. Also, it was possible to early detection of DUMP carrier by using PCR-RFLP(AvaⅠ). As the results of detection for BLAD(bovine leukocyte adhesion deficiency), BLAD carrier was not found in 16 proven bulls. But 5 candidtae bulls are discriminated to BLAD carrier, and it could be predicted to transmitted pathway of inherited loci by pedigree identification. Also, when digesting PCR products using restriction enzyme, results from TaqⅠ restriction enzyme were more efficient than that of HaeⅢ. After detection test of citrullinaemia, it was concluded that proven and candidate bulls were not. However, wide range of research and citrullinaemia genotyping should be performed. As a result of this study, the wide and various research should be performed in genetic disease of animal. And in the selection and breeding of animal, the breeding scheme by completely and continuously management of pedigree should be established.
The effects of exogenous fibrolytic enzymes (EFE; a mixture of two preparations from Trichoderma spp., with predominant xylanase and ${\beta}$-glucanase activities, respectively) on colonization and digestion of ground barley straw and alfalfa hay by Fibrobacter succinogenes S85 and Ruminococcus flavefaciens FD1 were studied in vitro. The two levels (28 and 280 ${\mu}g$/ml) of EFE tested and both bacteria were effective at digesting NDF of hay and straw. With both substrates, more NDF hydrolysis (p<0.01) was achieved with EFE alone at 280 than at 28 ${\mu}g$/ml. A synergistic effect (p<0.01) of F. succinogenes S85 and EFE on straw digestion was observed at 28 but not 280 ${\mu}g$/ml of EFE. Strain R. flavefaciens FD1 digested more (p<0.01) hay and straw with higher EFE than with lower or no EFE, but the effect was additive rather than synergistic. Included in the incubation medium, EFE showed potential to improve fibre digestion by cellulolytic ruminal bacteria. In a second batch culture experiment using mixed rumen microbes, DM disappearance (DMD), gas production and incorporation of $^{15}N$ into particle-associated microbial N ($^{15}N$-PAMN) were higher (p<0.001) with ammoniated (5% w/w; AS) than with native (S) ground barley straw. Application of EFE to the straws increased (p<0.001) DMD and gas production at 4 and 12 h, but not at 48 h of the incubation. EFE applied onto S increased (p<0.01) $^{15}N$-PAMN at 4 h only, but EFE on AS increased (p<0.001) $^{15}N$-PAMN at all time points. Prehydrolysis increased (p<0.01) DMD from both S and AS at 4 and 12 h, but reduced (p<0.01) $^{15}N$-PAMN in the early stage (4 h) of the incubation, as compared to non-prehydrolyzed samples. Application of EFE to barley straw increased rumen bacterial colonization of the substrate, but excessive hydrolytic action of EFE prior to incubation decreased it.
Chaudhary, Ajay K;Chaudhary, Shruti;Ghosh, Kanjaksha;Shanmukaiah, Chandrakala;Nadkarni, Anita H
Asian Pacific Journal of Cancer Prevention
/
v.17
no.3
/
pp.1519-1529
/
2016
Background: Matrix metalloproteinase -2 (gelatinase-A, Mr 72,000 type IV collagenase, MMP-2) and -9 (gelatinase-B, Mr 92,000 type IV collagenase, MMP-9) are key molecules that play roles in tumor growth, invasion, tissue remodeling, metastasis and stem-cell regulation by digesting extracellular matrix barriers. MMP-2 and -9 are well known to impact on solid cancer susceptibility, whereas, in hematological malignancies, a paucity of data is available to resolve the function of these regulatory molecules in bone marrow mononuclear cells (BM-MNCs) and stromal cells of myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). Objectives: The present study aimed to investigate mRNA expression and gelatinase A and B secretion from BM-MNCs in vitro and genotypic associations of MMP-2 (-1306 C/T; rs243865), MMP-9 (-1562 C/T; rs3918242), tissue inhibitor of metalloproteinase -1 (TIMP-1) (372T/C; rs4898, Exon 5) and TIMP-2 (-418G/C; rs8179090) in MDS and AML. Results: The study covered cases of confirmed MDS (n=50), AML (n=32) and healthy controls (n=110). MMP-9 mRNA expression revealed 2 fold increased expression in MDS-RAEB II and 2.5 fold in AML M-4 (60-70% blasts). Secretion of gelatinase-B also revealed the MMP-9 mRNA expression and ELISA data also supported these data. We noted that those patients having more blast crises presented with more secretion of MMP-9 and its mRNA expression. In contrast MMP-9 (-1562 C/T) showed significant polymorphic associations in MDS (p<0.02) and AML (p<0.02). MMP-9 mRNA expression of C/T and T/T genotypes were 1.5 and 2.5 fold increased in MDS and AML respectively. In AML, MMP-2 C/T and T/T genotypes showed 2.0 fold mRNA expression. Only MMP-9 (-1306 C/T) showed significant 4 fold (p<0.001) increased risk with chemical and x-ray exposed MDS, while tobacco and cigarette smokers have 3 fold (p<0.04) risk in AML. Conclusions: In view of our results, MMP-9 revealed synergistic secretion and expression in blast crises of MDS and AML with 'gene' polymorphic effects and is significantly associated with increased risk with tobacco, cigarette and environmental exposure. Release and secretion of these enzymes may influence hematopoietic cell behavior and may be important in the clinical point of view. It may offer valuable tools for diagnosis and prognosis, as well as possible targets for the treatments.
Twenty-two Phellinus strains were characterized using colony morphologies and polymerase chain reaction (PCR) to divide into Phellinus linteus. There were some differences in mycelial growth and colony shapes among the strains when they were grown on various media such as PDA, MCM, MEA and YM. Phellinus linteus was slowly growing, formed golden-yellow colony, and produced blue pigment on PDA media. When the regions of internal transcribed spacer (ITS) were amplified from ribosomal RNA (rRNA) coding genes of P. igniarius and P. linteus strains by means of PCR, two types of band (700 bp and 800 bp) were appeared, respectively. For the amplified intergenic region I (IGRI), P. igniarius strains showed a different band among 500, 600, 700 and 800 bp according to the strains, whereas P. linteus strains did one specific band of 700 bp. By polymorphism analysis after digesting the amplified products with 6 different restriction enzymes, a band specific to P. linteus was generated when the products for ITS region were digested with HaeIII, suggesting that the enzyme digestion could provide effective method to distinguish between P. igniarius and P. linteus. And also, the analysis of genetic relationship showed that the genetic similarities were 89% and 95% in P. igniarius and P. linteus strains, respectively. Random amplification polymorphic DNA (RAPD) analysis using multiple primer sets and arbitrarily primed PCR (AP-PCR) with ITS3 primer could also result in a reproducible way to identify P. linteus strains.
In this study, a dry single-phase anaerobic digestion process (Dranco system) was investigated to evaluate the optimum operational conditions. Several factors such as injection rate of organic waste, biogas production, $CH_4$ content in the biogas, pH of the sludge, $NH_3$-N and VFA concentration were investigated based on the operation of the digestion process for 2 months (short term) and 8 months (long-term). The operation results showed that a small quantity of food waste should be injected every week and that a 10% increase of the microorganism injection rate should be needed. However, normal operation was conducted after 11 weeks based on the designed quantity. The $CH_4$ content in the biogas was high at the beginning and the end of the food injection. However, it was low during week days. When the biogas production was high, the $CH_4$ concentration was low. The biogas production increased with an increase of the injection rate. $100m^3$/ton of biogas was produced from normal operation of the digestion process based on the designed quantity. The pH values of the digestion tank based on short-term operation ranged from 8 to 8.5. However, the pH values ranged from 7.45 to 8.15 after 4 weeks of long-term operation. The $NH_3$-N concentration of short-term operation ranged from 4,500 to 5,500 ppm and it gradually decreased to 2,000ppm after normal operation was commenced. For long-term operation, it was 5,000ppm initially and 3,800ppm after normal operation was commenced. The VFA concentration of sludge was less than 900ppm and 2,500ppm for short and long-term operations, respectively, after normal operation. Overall, the differences between sludge pH, $NH_3$-N and VFA concentrations may be due to the different types of microorganisms and the digestion ability of the microorganisms which exist in the accumulation of non digested organics. Moreover, it may be also caused by the type of food waste. Further investigation is needed to confirm these relationships.
We have previously developed three stage methane fermentation system capable of digesting food wastes effectively and then releasing high organic wastewater as a final product. In this study, we tried to devise an ecological water treatment system, which can efficiently remove the nitrogen and phosphorus contained in the organic wastewater. The system was made of microbiological filters, algae, and waterfleas. Of two species of alga tested, Selenastrum capricornutum showed higher growth rate and more efficiently removed the nitrogen from the wastewater than by Chlorella sp. In addition, the highest growth rate and the nitrogen removal efficiency could be obtained when high concentrations of $Mg^{2+}\; and\; Ca^{2+}$ were added to the diluted wastewater and the molar ratio of nitrogen to phosphorus was adjusted to 10 : 1. In this study the population relationship between alga and water flea was also examined in a test tube. The initial number of algal cells decreased as the waterflea population increased. However, the number of algal cells gradually increased again when waterflea population decreased partly due to the environmental resistance. From these results, it was believed that the ecological water treatment system could be used for removing the nitrogen and phosphorus from organic wastewater very effectively. Moreover, the waterflea cultured by this system as a final predator could be used as a good foodstuff for fishes.
A raw starch saccharifying enzyme from Aspergillus sp. SN-871 was purified by ammonium sulfate precipitation, DEAE-cellulose column chromatography, CM-Sephadex C-50 column chromatography and Sephadex G-75 gel filtration. The specific activity of purified enzyme was 18 fold and the yeild was 13.40%. The molecular weight of the purified enzyme was estimated as approximately 40,000 dalton by the method of Andrews gel filtration. The optimum pH and temperature for this enzyme were found to be 4 and $40^{\circ}C$, respectively and the stable range of pH was 2 to 5. The enzyme was themostable at below $60^{\circ}C$ and inactivated at $70^{\circ}C$. It showed a tendency to increase the enzyme activity under the presence of 0.01 M $BaCl_2$, but under 0.01 M$Pb(NO_3)_2$, $AgSO_4$, and $K_3Fe(CN)_6$ and citric acid etc. inhibited it completely. The substrate specifity of enzyme showed a tendency to increase the enzyme activity under addition of dextrin and glycogen, but under saccharose inhibited it. COD removal rate of Aspergillus sp. SN-871 was approximately 67 to 68%.
In grape vineyards, whitish spots in a cloud shape have been often observed on the fruit surface recently. However, the cause of the halo spot symptom was unknown, hindering countermeasures to be properly designed for the control. A small hole in the middle of the formless halo spot remained as a scar formed by oviposition of the thrips. It became later a suberized scab, which is separated from the epidermal cells on the surface either to be retained on or to be detached from it as time proceeds. Such a symptom is distinguished from the feeding damages caused by thrips or true bugs occurring on the grape fruits. With DNA extracted from the egg-shell found in the hole, molecular diagnosis by amplifying an ITS2 region with universal primers and subsequently digesting the PCR product by an restriction enzyme (RsaI) revealed that the egg was laid by Frankliniella occidentalis. In addition, a mitochondrial COI sequence confirmed that the halo spot symptom was formed by its oviposition. This study provides accurate information on the peculiar damage symptom caused by oviposition of F. occidentalis that could be useful in the control strategies for this pest in vineyards.
About 500 bacterial and fungal strains from a wide variety of natural habitats were screened for a new type II restriction endonuclease. Among the 500 species, we selected one species that produced a new restriction endonuclease. This strain has an optimum temperature of $30^{circ}C$ for growth. Morphological, cultural, and physiological characteristics were examined for identification of the isolated strain J-482. This strain was found to belong to the genus Alcaligenes. The restriction endonuclease was named as AspJI and partially purified from Alcaligenes sp. J-482 by DEAE-Sephadex A-50 column chromatography and gel filtration. Most of other nucleases were removed by the purification steps. The AspJI has a substrate specificity to ${lambda}$ DNA, pBR322 and Adenovirus-2 DNA. For its maximal activity, the isolated enzyme requires $MgCl_2$, which should be at least 12.5 mM and it does not need any other cofactors. It is maximally active in the absence of NaCl and is completely inactivated at 100 mM NaCl. The pH and temperature optima for activity were pH 7.5 and $37^{circ}C$, respectively. The DNA fragments generated by digesting ${lambda}$ DNA, pBR322, and Adenovirus-2 DNA with AspJI were the same as that produced by AatII. This suggests that AspJI is an isoschizomer of AatII.
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