unidad_COVID2019
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- Mar 2020
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The new virus is genetically 96% identical to a known coronavirus in bats and 86-92% identical to a coronavirus in pangolin. Therefore, the transmission of a mutated virus from animals to humans is the most likely cause of the appearance of the new virus.
Source of COVID-19
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medium.com medium.com
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When asked why Wuhan was so much higher than the national level, the Chinese official replied that it was for lack of resources, citing as an example that there were only 110 critical care beds in the three designated hospitals where most of the cases were sent.
Wuhan's rate then = 4.9%
National rate = 2.1%
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80% of food in the UK is imported. How about your country? Your town?
Food for thought
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The South Korean government is delivering food parcels to those in quarantine. Our national and local governments need to quickly organise the capacity and resources required to do this.Japanese schools are scheduled to be closed for march.
Food delivery in South Korea and closing schools in Japan
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The limited availability of beds in Wuhan raised their mortality rate from 0.16% to 4.9%This is why the Chinese government built a hospital in a week. Are our governments capable of doing the same?
Case of Wuhan
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The UK population is 67 million people, that’s 5.4 million infected.Currents predictions are that 80% of the cases will be mild.If 20% of those people require hospitalization for 3–6 weeks?That’s 1,086,176 People.Do you know how many beds the NHS has?140,000
There will be a great lack of beds
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Evolving to be observant of direct dangers to ourselves seems to have left us terrible at predicting second and third-order effects of events.When worrying about earthquakes we think first of how many people will die from collapsing buildings and falling rubble. Do we think of how many will die due to destroyed hospitals?
Thinking of second and third-order effects of events
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Can you guess the number of people that have contracted the flu this year that needed hospitalisation in the US? 0.9%
0.9% of flu cases that required hospitalisation vs 20% of COVID-19
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The UK has 2.8 million people over the age of 85.The US has 12.6 million people over the Age of 80.Trump told people not to worry because 60,000 people a year die of the flu. If just 25% of the US over 80’s cohort get infected, given current mortality rates that’s 466,200 deaths in that age group alone with the assumption that the healthcare system has the capacity to handle all of the infected.
Interesting calculation of probabilistic deaths of people > 80. Basically, with at least 25% infected people in the US who are > 80, we might have almost x8 more deaths than by flu
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The formula China is using is current total deaths/current confirmed cases.
Mortality rate of COVID-19 used by China =
\(\dfrac{currentTotalDeaths}{currentConfirmedCases}\)
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Each year the flu infects between 2 to 11% of the population. 11The infectiousness of a disease is measured by its R0 value. R0 is a measure of how many people will catch a disease from one infected person. The R0 value of the flu is 1.28. The R0 of COVID-19 is assumed to be between 1.4–3.8
R0 - measure of how many people will catch a disease from one infected person.
R0 (Flu): 1.28
R0 (COVID-19): 1.4 - 3.8 (but some say 4.7 - 7.12)
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cases outside of china are doubling roughly every 5 days. The reported rate of infection in China is lower, but China has taken drastic quarantine measures, including welding people inside apartment buildings.
Doubling tendency of COVID-19
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“confirmed cases” is the lowest possible number of cases in that country. It’s the lower bound of an estimate.
What confirmed cases actually means
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In The Atlantic, Marc Lipsitch, a leading epidemiologist at Harvard reported that “that within the coming year, some 40 to 70 percent of people around the world will be infected with the virus that causes COVID-19”. This was last week and while ridiculed at the time, his assumptions are now the generally accepted position among epidemiologists.
40 - 70 % of people will be infected with COVID-19 (it might even increase)
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Death Rate
Death rate of COVID-19:
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The Virus may persist on surfaces for up to 9 days, on some surfaces up to 27 days
Coronavirus' lifespan
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- Jan 2020
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www.ncbi.nlm.nih.gov www.ncbi.nlm.nih.gov
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RRID:ZFIN_ZDB-ALT-170522-19
DOI: 10.7554/eLife.42881
Resource: (ZFIN Cat# ZDB-ALT-170522-19,RRID:ZFIN_ZDB-ALT-170522-19)
Curator: @evieth
SciCrunch record: RRID:ZFIN_ZDB-ALT-170522-19
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- Aug 2019
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www.americanyawp.com www.americanyawp.com
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Say howdy to George Carter, and thank him for taking the pistol from you when you were shooting at me. From your old servant,
Great closing
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I am doing tolerably well here. I get twenty-five dollars a month, with victuals and clothing; have a comfortable home for Mandy,—the folks call her Mrs. Anderson,—and the children—Milly, Jane, and Grundy—go to school and are learning well. The teacher says Grundy has a head for a preacher. They go to Sunday school, and Mandy and me attend church regularly. We are kindly treated.
Civil War and emancipation change Jourdan’s life and The life of his wife and children.
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- Jun 2019
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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IL-2 Bioassay
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IL-2 Bioassay
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IL-2 Bioassay
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Assay of IFN-yand IL-10 using sandwich ELISA
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IL-2 Bioassay
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LympllOkine Assays
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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RNA extraction and cDNA synthesis
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Transblotting
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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AnalysisofAP-landNFKBactivatio
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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Descriptive Statistics
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- May 2019
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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The CLD-J domain shares ~51 % similarity with the CDPK from Arabidopsis thaliana AtCPK-l. The homology model of CLD-JD was determined using Swiss Model from EMBL. The template model used was CLD-JD of AtCPK-1, which was crystallized as a dimer. The J -domain helices from the two monomers were swapped with each other in this structure (Chandran et aI., 2006). Therefore, the initial homology model generated for the complementary CLD-J domain for PfCDPK4 was also a dimer. To understand the interaction of this helix (Gln358_ Lys371) with CLP of the monomer, this helix was rotated and translated keeping residues 372-375 as the flexible linker region and superimposed on to the helix from the other monomer, which resulted in the initial model for the CLD-J domain monomer. Initially, these flexible linker residues (372-375) were locally minimized using COOT (Emsley and Cowtan, 2004), and the overall structure was refined with slow cooling using annealing of CNS (Brunger et aI., 1998) to remove all the short contacts. Finally, the model quality was checked with the Pro check software (Laskowski et aI., 1996). The homology model was generated with the help of Dr. S. Gaurinath, JNU
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Homology Modeling
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sg.inflibnet.ac.in sg.inflibnet.ac.in
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released from the para-ovarian pad of fat as well as from the peritoneal reflections while care was taken to avoid injury to the ovarian vessels. A ligature was tied around the distal end of the fallopian tube including the ovarian vessels following which the ovary was excised. Hemostasis was secured before the stump of the tube was pushed back into the peritoneal cavity. The peritoneum was closed by continuous sutures using 2-0 silk. The same protocol was followed to perform oophorectomy on the contralateral side. The muscular layer and skin were closed together using surgical clips
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The anesthetized mice were operated under strict aseptic conditions inside a laminar flow hood. The mouse was placed over layers of sterile tissue paper and the skin overlying the dorsal flanks was sterilized by wiping with 70% ethanol. The flank was palpated gently to identify the kidney, and an incision(~ 5 mm) was made using a pair of scissors on the overlying skin which penetrated the skin, sub-cutaneous tissue and the muscle layer with the parietal peritoneum being exposed and intact. The para-ovarian pad of fat was identified through the intact peritoneum and a small incision was made on the peritoneum overlying it. The ovarian tissue along with the fallopian tube was mobilized and delivered through the incision site. The ovary was
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Bilateral oophorectomy
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Hair from the skin overlying the left and right dorsal flanks were removed using electrically operated razor. The skin overlying the abdomen was sterilized by wiping with 70% ethanol. Ketamine (1 00 mg/kg) and xylocaine (2%) (20 mg/kg) were mixed and administered intraperitoneally. The mice were returned to the cage and the onset of anesthetic effect was monitored. The mice were considered to be in surgical anesthesia when there was loss of palpebral reflex, righting reflex, and toe pinch reflex. Respiratory rate and heart rate were monitored continuously.
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General anesthesia:
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Bilateral oophorectomy, the surgical removal of both the ovaries, was performed in mice to simulate a condition of estrogen depletion. All procedures in mice were performed after obtaining approval from the Institutional Animal Ethics Committee (National Institute of Immunology, New Delhi). Female BALB/c mice were used in the study
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Bilateral oophorectomy and sham surgery in mice
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Animal experiments
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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in IOmM Tris/HCI (pH 7.4)/ lmM EDTN 0.5% SDS. The radioactivity associated with the cells was counted in a y-counter (LKB).
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Competition binding analysis was performed to compare the affinity of the chimeric toxins with native antibody. Anti-transferrin receptor antibody (HB21) was iodinated using iodogen method as described by Harlow and Lane, (1988). Adherent cell lines, namely, A431 and A549 were plated at 4Xl05 cells per well in a 24 well plate and used 16 h later for the assay. 500 J.Ll medium, same as described in cytotoxicity assay, was used to dispense the cells. HUT102 cells were also plated at the same density in microfuge tubes and used immediately. After 2 washes with binding buffer (0.1% BSA in DMEM), various dilutions of toxin, along with 3 ng of labeled antibody in binding buffer, were added to the cells. The cells were incubated at 25 °C for 2 h with mild shaking, washed three times with binding buffer and lysed
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Binding Studies
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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then estimated by monitoring its competitive binding to the hCG recepto~ in the presence of radiolabelled hCG. The testes homogenates from 10 -14 week old Wistar outbred rats were prepared by the method of Dighe and Moudgal ( 1983 ), and the receptor assay 1975 ), with 1989 ) .
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Culture supernate from stable clones secreting alpha hCG was mixed with the culture supernate from stable clones showing J3hCG activity. The mixing was done with shaking at 37°C for 16 hours, to allow the two subunits to associate. The presence of the heterodimer in the culture supernate was
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Radioreceptor assay.
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shodhganga.inflibnet.ac.in shodhganga.inflibnet.ac.in
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EROD
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ITALYusinga96wellmicrolitreplatereadbyELISAmicroplatereader(modelEL3/Sx,BioTeKInstrumentsINC).Thefinalsolutionwasreadatawavelengthof450nm.Theplasmacortisolconcentrationwascalculatedbasedonaseriesofstandards.
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PlasmacortisolwasmeasuredbyadirectimmunoenzymaticdeterminationofcortisolkitmanufacturedbyEquiparSriviaG.Ferrari,21/N-21047,SARONNO
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Plasmacortisol
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Thecapabilityofheadkidneyneutrophilstomovewasassayedbyamigration-under-agarosetechniquemodifiedfromNelsonetal.(1975).ThemethodhasbeendescribedbySaloetal.(1998).Thedistance,thecellshadmigratedfromthemarginofthewelltowardsthewellcontainingcasein(directedmigration)andintheopposite direction(randommigration)weremeasuredunderthemicroscope.
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Migration
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Phagocytesfromtheheadkidneywerestimulatedwithphorbol12-myristate13 -acetate(PMA,SigmachemicalCo)andtherespiratoryburstwasdeterminedbytheluminol-enhancedCLmethod(ScottandKlesius,1981)
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Theenzyme-linkedimmunospot(ELISPOT)assaywasusedfortheenumeration of totalimmunoglobulinsecretingcells(ISC)andantigen(BCG)-specificantibody-secretingcells(ASC)inthespleenandtheblood.TheELISPOTassayfollowedby Aaltonenetal.(1994)wasused.
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EnumerationofsecretinglymphocyteswithELISPOTassay
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TheamountoftotalIgMandspecificantibovineY-globulin(BGG)antibodyinthefishplasmawasmeasuredbyanenzyme-linkedimmunosorbentassay(ELISA)asgiven byAaltonenetal.(1994)
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Serumimmunoglobulin
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Theheadkidneywasusedasasourceofphagocytesforchemiluminescence (CL)andmigrationassays.Cellsfromthehomogenizedheadkidneywereseparatedwithatwo-stepPercollgradient.Granulocyteswerecollectedatthe1.070-1.090g/cm3interfaceandafterwashingwithrHBSSresuspendedinphenolred-freerRPMl.
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Headkidney
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Thespleenswereremovedandhomogenisedindividuallythroughanylonnet(80mesh).Forisolationoflymphocytes,thetissuehomogenatewaslayeredonatwo-stepPercolldensitygradientandcentrifugedfor30minat400xg.Thelymphocyteswerecollectedatthe1.040-1.080g/cm3interface,washedtwice(400xg,10min)withrHbss,andresuspendedin2mlrRPMl.Cellswerecountedbytrypanblueexclusioninahaemocytometer(viability>95%)andthenumberoflymphocytesfrombloodandspleenwereadjustedto2x106/ml
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Spleen
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Abloodsamplewastakenfromthecaudalveinofeachfishwitha1mlheparinisedsyringeand24-gaugeneedle.Thebloodwascentrifuged(400xg,5min)fortheseparationofplasma.Plasmawasstoredfrozen(-70°C)forthedeterminationoftheimmunologicalparameters
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Collectionofplasma
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Immunologicalanalysis
Tags
- Method-19-Method-9
- Method-19-Method-8
- Method-19-Method-5-detail
- Method-19-Method-3
- Method-19-Method-4-detail
- Method-19-Method-2-detail
- Method-19-Method-1
- Method-19-Method-1-detail
- Method-19-Method-6
- Method-19
- Method-19-Method-5
- Method-19-Method-7-detail
- Method-19-Method-7
- Method-19-Method-8-detail
- Method-19-Method-4
- Method-19-Method-3-detail
- Method-19-Method-2
Annotators
URL
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