4,556 Matching Annotations
  1. Jul 2019
  2. sg.inflibnet.ac.in sg.inflibnet.ac.in
    1. Haemocyte morphogenesis
    2. Spreading inhibitory behavior
    3. Inhibition of haemocyte aggregation
    4. Haemolymph protein profile
    5. Total haemocyte count assay
    6. Immunomodulatory
    7. Helicoverpa armiger
    8. Spodoptera litura
    9. Gut enzyme profile
    10. Insecticidal activity
    11. . Statistical analysis
    12. Inhibition of haemocytes spreading behavior
    13. . Inhibition of haemocytes aggregation behavior
    14. Haemolymph protein profiling
    15. Total haemocyte count (THC)
    16. Haemolymph collection
    17. Immunomodulatory
    18. Lactate dehydrogenase
    19. Asparate (AAT) and Alanine aminotransferase (ALT)
    20. Gut enzyme profile
    21. Oral toxicity bioassay
    22. Microinjection bioassay
    23. VS preparation
    24. Insect collection
    25. INSECTICIDAL AND IMMUNOMODULATORY ACTIVITY AGAINST INSECT PEST
  3. Jun 2019
    1. Increased Chemokine and Toll like receptor on T cells after vaccination in HBV positive newborns.
    2. Cord Blood immune profiles of HBV positive newborns at birth(Cord blood vs. peripheral blood)
    3. CD107a expression (marker of cytotoxicity)
    1. The a-PMB chain was subjected to acid-acetone treatment to separate the heme from the a globin. Briefly, a solution of concentrated a-PMB chain (5 ml; 30 mg/ml) was added dropwise to I 00 ml of thoroughly chilled acid-acetone solution (0.5% v/v HCI in acetone) with constant shaking, and then incubated at -20°C for 30 min to allow complete precipitation of the globin. The precipitated globin was isolated by centrifugation at 7000 rpm (4°C) for 15 min and the supernatant containing soluble heme was discarded
    2. Preparation of heme-free a chain
  4. May 2019
    1. enginethatistheproblembut,rather,theusersofsearchengineswhoare.Itsuggeststhatwhatismostpopularissimplywhatrisestothetopofthesearchpile
      • I wanted to highlight the previous sentence as well, but for some reason it wouldn't let me*

      I understand why the author is troubled by the campaign's opinion of "It's not the search engines fault". It makes it seem as if there was nothing that could be done to stop promoting those ideas, and that if something is popular it will just have to be the result at the top.

      This can be problematic, as people who were not initially searching that specific phrase may click through to read racist, sexist, homophobic, or biased information (to just name a few) that perpetuates inaccuracies and negative stereotypes. It provides easier access into dangerous thinking built on the foundations of racism, sexism, etc.

      If the algorithms are changed or monitored to remove those negative searches, the people exposed to those ideas would decrease, which could help tear down the extreme communities that can build up from them.

      While I do understand this view, I also think that system can be helpful too. All the search engine does is reflect the most popular searches, and if negative ideals are what people are searching, then we can become aware and direct their paths to more educational and unbiased sources. It could be interesting to see what would happen if someone clicked on a link that said "Women belong in the kitchen", that led them to results that spoke about equality and feminism.

    1. 'Sam!' he called. 'Pippin! Merry! Come along! Why don't you keep up?'10There was no answer. Fear took him, and he ran back. As he struggled on he called again, and kept on calling more and more frantically. He was weary, sweating and yet chilled. It was wholly dark.'Where are you?' he cried out miserably.There was no reply. He stood listening. He was suddenly aware that it was getting very cold, and that up here a wind was beginning to blow, an icy wind. A change was coming in the weather. The mist was flowing past him in shreds and tatters. His breath was smok­ing.11 He looked up and saw with surprise that faint stars were ap­pearing overhead amid the strands of hurrying cloud and fog. Oat of the east the biting wind was blowing.'Where are you?' he cried again, both angry and afraid.'Here!' said a voice, deep and cold, that seemed to come out of the ground. 'I am waiting for you!''No!' said Frodo; but he did not run away. His knees gave,12 and he fell on the ground. Nothing happened, and there was no sound. Trembling he looked up in time to see a tall dark figure like a shadow against the stars. It leaned over him. He thought there were two eyes, very cold though lit with a pale light that seemed to come from some remote distance. Then a grip stronger and colder than iron seized him. The icy touch froze his bones, and he remembered no more.When he came to himself again, for a moment he could recall nothing except a sense of dread. Then suddenly he knew that he was imprisoned, caught hopelessly; he was in a barrow. A Barrow-wight had taken him, and he was probably already under the dreadful spells of the Barrow-wights about which whispered tales spoke. Hedared not move, but lay as he found himself: flat on his back upon a cold stone with his hands on his breast.As he lay there, thinking and getting a hold on himself, he no­ticed all at once that the darkness was slowly giving way:13 a pale greenish light was growing round him. He turned, and there in the cold glow he saw lying beside him Sam, Pippin, and Merry.There was a loud rumbling sound, as of stones rolling and fal­ling, and suddenly light streamed in. A low door-like opening appeared at the end of the chamber beyond Frodo's feet; and there was Tom's head against the light of the sun rising red behind him.'Come, friend Frodo!' said Tom. 'Let us get out on to the clean grass! You must help me bear them.' Together they carried out Merry, Pippin and Sam. To Frodo's great joy the hobbits stirred, robbed their eyes, and then suddenly sprang up. They looked about in amazement. 'What in the name of wonder?14 began Merry. 'Where did you get to, Frodo?''I thought that I was lost', said Frodo; 'but I don't want to speak of it.' But Tom shook his head, saying: 'Be glad, my merry friends, and let the warm sunlight heat now heart and limb! Cast off these cold rags! Run naked on the grass!'
    1. A stock solution of xylose (1 mg mL-1) was prepared in distilled water. A dilution series ranging from 100-1000 μg mL-1 was prepared from the stock solution. To 1 mL of solution, 1mL of DNSA was added and kept in a boiling water bath for 10 min and then 400 μL of sodium potassium tartrate solution was added and kept it for cooling. The absorbance was recorded in a spectrophotometer (Shimadzu, UV-VIS) at 540 nm
    2. Preparation of standard curve of xylose
    3. Transformation of calcium-competent cells was carried out by the procedure detailed below: •The competent bacterial cells were thawed briefly and 200 μL of cells was mixed rapidly with plasmid DNA (10-50 ng) in fresh, sterile microcentrifuge tubes and maintained on ice for 30 min. A negative control with competent cells only (no added DNA) was also included. •Cell membranes were disrupted by subjecting cells to heat-pulse (42 °C) for 90 sec. •After heat shock, cells were incubated on ice for 5 min. •Cells were then mixed with 1 mL LB medium and incubated with shaking at 37 °C for 1 h. •For blue/white screening 40 μL of X-gal solution (20 mg mL-1 in dimethylformamide) and 4 μL of the IPTG (200 mg mL-1) was spread on LB-ampicillin (LB-amp) plates with a sterile glass rod. The plate was allowed to dry for 1h at 37 °C prior to spreading of bacterial cells. •Bacterial cells (100-200 μL) were spread and the plate was incubated at 37 °C for overnight. •White colonies were picked from the plates and suspended into LB-amp broth and cultivated to OD600=0.5
    4. Transformation procedure
    5. PurifiedDNA fragments of size 2-8 kb were ligated to the treated vector using a 1:3::vector :insert ratio in a volume of 10 μL. The total amount of DNA was about 0.5 μg. Vector and insert DNA was heated to 45 °C for 10 min and the immediately chilled on ice for 5 min prior to addition of ligase and buffer. T4 DNA ligase (NEB, England) was added to a final concentration of 0.125 UμL-1 and reactions were incubated at 16 °C for overnight in a ligation chamber. Reaction mixture incubated under same condition without addition of the enzyme was used as control. A ligation reaction was also set up under condition with linear plasmid DNA containing the
    6. Ligation of insert DNA with dephosphorylated vector
    7. In order to minimize self ligation of vector during cloning experiments, the digested DNA was subsequently treated with calf intestinal phosphatase (CIP) [NEB, UK]. The reaction conditions and amount of CIP were optimized and varied from (0.06-1) unit/picomole DNA termini. The dephosphorylation reaction was carried out in 50 μL reaction as follows. Reaction mixture containing no restriction enzyme was treated as control. Reaction was incubated for 1 h at 37 °C and stopped by heat inactivation at 65 °C for 20 min. 2.5.5. Composition of restriction mixture (50 μL) Linearized Plasmid DNA X μL (1 μg) CIP 1 μL (0.06-1 U μL-1) Reaction buffer (10X) 5.0 μL Distilled water Y μL Total volume 50 μL Linearized and dephosphorylated plasmids from each reaction were purified from low melting agarose gel using gel extraction method according to the manufacturer’s protocol (Qiagen gel extraction kit, Germany). 100 ng DNA from each reaction was then ligated in15 μL reaction volume containing 1.5 μL of 10X ligation buffer (NEB, England) and 0.2 μL of T4 DNA ligase to check the efficiency of self ligation after dephosphoryaltion. The ligation mixture was incubated at 16 °C for overnight and transformed into E. coli DH5αcompetent cells.
    8. Dephosphorylation of the restricted plasmid
    9. The vector isolated as above was digested with BamHI to generate the cohesive ends. The reaction was performed in 1.5 mL Eppendorf tubes as described below. Composition of restriction mixture (100 μL) Plasmid DNA X μL (20 μg) Bam HI 8 μL (10 U μL-1) NEB buffer 4 10.0 μL BSA (100X) 1 μL MQ water Y μL The reaction mixture was incubated at 37 °C for 3 h. The digestion was stopped by heat inactivation at 65 °C for 20 min. The digestion of plasmid was checked using 1.2 % (w/v) agarose gel electrophoresis for linearization of the plasmid. The digested plasmid was purified from low melting agarose gel using gel extraction method according to the manufacturer’s protocol (Qiagen gel extraction kit, Germany).
    10. Restriction digestion of plasmid DNA
    11. Two hundred μL of alkaline-SDS solution was added to the above suspension, mixed by inverting the tubes up and down 3 times and incubated for 5 min at room temperature. ƒTo the above mixture, 250 μL of 3 M Na-acetate (pH 4.8) was added, mixed by inverting the tubes up and down 3 times, and centrifuged at 12,000 x g for 10 min. ƒThe supernatant was collected in another micro centrifuge tube (MCT), 200 μL of phenol:chloroform solution was added, inverted two times and centrifuged at 12, 000 x g for 8 min at room temperature. ƒThe aqueous phase was transferred to new tubes and 500 μL of chilled (-20 °C) ethanol (96 %) was added. ƒThe tubes were centrifuged at 13,000 x g for 25 min at 4 °C, supernatant discarded and pellet dried for 15 min at room temperature. ƒThe pellet was washed with 500 μL of chilled 70 % (v/v) ethanol and centrifuged at 13, 000 rpm for 4 min at 4 °C. ƒThe pellet was dried at room temperature and dissolved in 50 μL of 1X TE buffer (pH 8.0) containing RNase and stored at -20 °C till further use.
    12. The cells of E. coli DH10B having p18GFP vector were cultivated for overnight at 37 °C in LB medium containing ampicillin (100 μg mL-1). ƒThe E. coli culture having p18 GFP vector (~1.5 mL) was taken in Eppendorf tubes and centrifuged at 10, 000 x g for 5 min. ƒThe pellet was homogenized by vortex mixing in 100 μL of homogenizing solution
    13. Plasmid isolation from miniprep method
    14. The metagenomic DNA extracted from above defined protocol was digested with Sau3A1 at conditions optimized to generate maximum fragment in the size range of 2-6 kb. Different concentration (0.05 to 1 unit) of enzyme was used to optimize the digestion of 1 μg of DNA. Reactions were carried out in a final volume of 30 μl each in an Eppendorf of 1.5 mL. Reaction mixture (1 μg DNA having 3 μL NEB buffer 3 and 0.3 μL of 10X BSA) were kept at 37 °C for 10 min and stopped by heat inactivation at 80 °C for 20 min. Different digested reactions were checked for the desired fragments using 0.8 % (w/v) agarose gel electrophoresis. After optimization of DNA fragments for the appropriate size, a large scale digestion was carried out and the fragments (2-8 kb) were purified from low melting agarose gel using gel extraction method according to the manufacturer’s protocol (Qiagen gel extraction kit, Germany)
    15. Insert DNA preparation
    16. CONSTRUCTION OF METAGENOMIC LIBRARY
    17. An attempt was made to study the effect of storage of DNA extracts on DNA yield and purity. The DNA extracts were centrifuged and the supernatants were dispensed into 2.0 mL Eppendorf tubes and stored at -20 oC for a month. DNA precipitation and its quantification were carried out at a week intervals.
    18. Effect of storage on soil/sediment DNA extracts
    19. The isolated DNA was diluted (1:100) with MQ. The concentration (mg mL-1) of the DNA [N] was determined spectrophotometrically by recording absorbance at 260 nm (A260) as: A260 = ε 260[N]where ε 260 is the extinction coefficient of DNA (50 for ds DNA) [N] = concentration (mg mL-1) of DNA The concentration of ds DNA [N] was calculated as [DNA] (mg mL-1) = A260/ε 260 [DNA] (μg mL-1) = A260 × 50 × dilution factor Purity of DNA was checked by measuring absorbance at 260 and 280 nm and calculating the A260/A280 ratio (Sambrook et al., 1989). A DNA sample was considered pure when A260/A280 ranged between 1.8-1.9. An A260/A280 < 1.7 indicated contamination of the DNA preparation with protein or aromatic substances such as phenol, while an A260/A230 < 2.0 indicated possible contamination of high molecular weight polyphenolic compounds like humic substances.
    20. Determination of DNA quantity and purity
    21. as well as commercial methods (MN kit, Germany; Mo-Bio kit, CA, USA; Zymo soil DNA kit, CA, USA) according to the manufacturer’s protocols and compared in terms of DNA yield and purity.
    22. The soil DNA from Pantnagar and Lonar soil samples were also extracted by various manual (Desai and Madamwar, 2007; Agarwal et al., 2001; Yamamoto et al., 1998
    23. Alternatively metagenomic DNA was extracted from the alkaline soil samples by using different commercial kits (UltraClean™, PowerSoil™ [Mo Bio Laboratories Inc., Carlsbad, CA, USA], Nucleospin kit [Macherey-Nagal, Germany] and Zymo soil DNA isolation kit [CA, USA]). The DNA was finally suspended in 100 μL of sterile Milli Q water for further analysis.
    24. Commercial kits