L'enseignant référent qui coordonne les équipes de suivi de la scolarisation est l'interlocuteur des familles pour la mise en place du projet personnalisé de scolarisation.

Instance methods Instances of Models are documents. Documents have many of their own built-in instance methods. We may also define our own custom document instance methods. // define a schema const animalSchema = new Schema({ name: String, type: String }, { // Assign a function to the "methods" object of our animalSchema through schema options. // By following this approach, there is no need to create a separate TS type to define the type of the instance functions. methods: { findSimilarTypes(cb) { return mongoose.model('Animal').find({ type: this.type }, cb); } } }); // Or, assign a function to the "methods" object of our animalSchema animalSchema.methods.findSimilarTypes = function(cb) { return mongoose.model('Animal').find({ type: this.type }, cb); }; Now all of our animal instances have a findSimilarTypes method available to them. const Animal = mongoose.model('Animal', animalSchema); const dog = new Animal({ type: 'dog' }); dog.findSimilarTypes((err, dogs) => { console.log(dogs); // woof }); Overwriting a default mongoose document method may lead to unpredictable results. See this for more details. The example above uses the Schema.methods object directly to save an instance method. You can also use the Schema.method() helper as described here. Do not declare methods using ES6 arrow functions (=>). Arrow functions explicitly prevent binding this, so your method will not have access to the document and the above examples will not work.

The first time the ratio of length to width was written in a letter dated 25 October 1786. This letter was from the German Georg Christoph Lichtenberg to Johann Beckmann. He wrote here about the advantages of basing paper on a √2 ratio. Lichtenberg is known for the ratio between length and width of a surface which remains the same after the narrated halving of the surface. The result is 1:√2.

Your comment inspires me to pay more attention to citing and clarifying my claims.

There are several occasions where the massebah is not associated with pagan worship. When the massebah is associated with the worship of Yahweh, the massebah is accepted as a valid expression of commitment to Yahweh.

in violation of the demands of the covenant, the people of Israel erected sacred stones dedicated to other gods (Hosea 10:1). In their religious reforms, both Hezekiah (2 Kings 18:4) and Josiah (2 Kings 23:14) destroyed the sacred pillars which the people of Israel had dedicated to the worship of Baal.

During the establishment of the covenant between Yahweh and Israel, the people were commanded to destroy the sacred stones of the Canaanites, “You must demolish them and break their sacred stones (masseboth) to pieces” (Exodus 23:24).

2 3-4 x 4 3-4 inches in size, made of seal grain , real sealor Russia leather, in a thoro

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Weread, for example, of Philistine incursions into the hill country, toMichmash in Benjamin (1 Samuel 13:23), and the Rephaim Valley nearJerusalem (2 Samuel 5:17–22). It was in one of these border disputes thatthe city at Khirbet Qeiyafa was conquered and destroyed.

hume is not in book one arguing that persons do not exist in fact in book two he's going to spend most of his time explaining what persons 01:17:41 are he when instead what he's claiming is that persons don't have selves

remind us of the historical reduction of the human to the status of ananimal under transatlantic slavery, but also were used as a mode of resistancefor enslaved peoples

5.1 Recognize that 1) the biggest threat to good decision making is harmful emotions, and 2) decision making is a two-step process (first learning and then deciding).

so let's suppose let's suppose your listeners are with me and you know we kind of agree like okay yes transformation's necessary and uh again i want to emphasize i'm not talking about reform i'm not talking 00:58:59 about a softer better capitalism i'm not talking about you know improved voter registration or like any of those things i'm talking about de novo starting over from scratch what might be 00:59:13 best and if it turns out that the old systems were better than anything that humanity can come up with well then you know that's the answer but i can't imagine that's true because the old systems were never designed in any kind of 00:59:25 you know thoughtful science driven [Music] you know process to to to test to explore and to come up with fitness like what is the you know we don't even have a fitness for our current society 00:59:39 much less of fitness for societal designs i mean we have the gdp but that's a terrible terrible limited fitness metric 00:59:51 okay so suppose you're with me suppose we're we're on board we we want to do this de novo design thing where do we start what's the what's what where do we even get off the 01:00:03 ground on this and i suggest that the way to do it is through first address worldview from world view once we understand what the world view is 01:00:15 what a reasonable useful world view will be for this project then then purpose derives worldview begets purpose once you understand what it is you want 01:00:28 what you value what do you value once you understand what you value then you can say well i value a and therefore the purpose is to 01:00:39 have a manifest in society for example so once you have purpose then you can think about what metrics how would you measure whether are you so 01:00:53 here's a new design is it fit for purpose does it do does it fulfill its purpose you know that's the question and then metrics go with some kind of fitness evaluation 01:01:05 and then finally last of all of those would be the design okay we know what we know what we value we know what this thing is supposed to do we know what the purpose is we know that attractor is supposed to you know plow the ground or something we 01:01:18 know what this is supposed to do we know how to measure success and uh now finally then let's talk about design what are the what are the you know the specifics and mechanics and 01:01:31 how does that happen and the the series is really kind of laid out this way the first paper really talks about world view and purpose the second paper talks about the you know the more the mechanics of things 01:01:44 like viability how would you make this thing viable things like that and then the very last paper that's titled the subtitle design okay so uh that's how we uh and 01:01:56 and maybe i will just mention here that i put metrics before design because we might have some ideas uh getting back to that preference factor we might have some ideas like we would like people not to die at 01:02:08 30 you know we'd like people to mostly live to a ripe old age and have you know enough water water to drink and food to eat and all that kind of stuff so uh you know what kind of design once 01:02:20 now that we have metrics to measure that kind of stuff longevity and nutrition and things what kind of designs would help us to reach those targets you know so that's one reason why design 01:02:31 why metrics comes before design okay

Matt Taibbi asked his subscribers in April. Since they were “now functionally my editor,” he was seeking their advice on potential reporting projects. One suggestion — that he write about Ibram X. Kendi and Robin DiAngelo — swiftly gave way to a long debate among readers over whether race was biological.

the combination of trading patterns and pref-erences of European planters in the Americas for laborers of specific Africanethnicities tended to lump together large numbers of captive Africans from cer-tain areas into particular colonies in the Americas.

L’esercizio fisico deve essere raccomandato per il controllo del diabete nelle persone con diabete di tipo 2?

help you save

Well, no. I oppose capital punishment, just as (in my view) any ethical person should oppose capital punishment. Not because innocent people might be executed (though that is an entirely foreseeable consequence) but because, if we allow for capital punishment, then what makes murder wrong isn't the fact that you killed someone, it's that you killed someone without the proper paperwork. And I refuse to accept that it's morally acceptable to kill someone just because you've been given permission to do so.

One solution that fixed this issue with my ISP was that when I went through the first and second line and got in touch with the people that fixed my problem, I asked them if they could give me one of their personal numbers in case the same problem happened again. The problem did occur a couple more times, and I just directly called the same guy.

Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

SUPPLEMENTARY DATA

SUPPLEMENTARY DATA

Source Data

Source Data

Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

Supplemental material

So unfortunately 1 step forward, 2 steps back.

Chapter 3Biopsychology

道地

真材實料

平價

清爽

滿嘴

推出

鳳梨酥

Sampling point, Sample Size and Sample collection

Sequence analysis

DNA sequencing of the 18S rDNA fragment

Purification of PCR product

Analysis of internal transcribed spacer region

RAPDand SSRscoring and data analysis

PCR amplification

Running of gel and visualization of DNA

Determination of the yield

Agarose gel electrophoresis

Qualitative and quantitative estimation of DNA

Determination of the yield

Procedure for DNA isolation

Reagents required for fungal DNA isolationand p

DNA isolation of Trichodermaisolate

Photography, evaluation and documentation

Procedurefor SDS-PAGE

Materialsrequired for SDS-PAGE

Protein profiling of bioagent through SDS-PAGE

Dinitrosalicylate reagent (DNS)(per liter)

Identificationof bioagent

Bacterial strains used

Production of Lipase

Antibiotic resistance amongVibriofrom shrimp farm

Extraction of genomic DNA

Validation of the predicted 3D structure

Phylogenetic and motif analysi

Phylogenetic and motif analysis of sequenced Dof domains

Gel elution of PCR products

Validations

Mapping of SbDof genes on sorghum chromosomes and its intron/exon gene structure prediction

Sequencing PCR

Minipreparation of plasmid DNA from transformed co

Restriction digestion

Basic requirements for PC

DNA extraction procedure

The table 6.1 gives the mixing probabilities and the associated parametricvalues fork(number of components) = 2,3, and 4. It may be noted thatthe Log likelihood value is smaller fork= 4 (the results fork= 5 , 6 etc.are not better than that fork= 4 and hence are not given here). The fourcomponents Poisson Mixture model is given in table 6.2. It may be notedthat 58% of wards may have higher incidence/relative risk and the remainingwards have lesser/lower incidence for the Cancer disease. We computed theposterior probability for each component for each ward (see table 6.3). Eachward is assigned to a particular component so that the posterior probability islarger. These results are also given in table 6.3 Finally we present Choroplethmaps based on those results

Algorithm

Data Sources

Poisson Model

Spodoptera litura

Haemolymph protein profiling

Venomous saliva optimization

Starvation and collection method

Insects Collection

Mandibular stylet

Head and stylet preparation

Growth maximum values

Rates of growth (OD678/day

Growth

Experimental Set up

Experiment 1: Assessing the impacts of elevated CO2and N levels on yield and nutrient uptake in rice

Temperature Gradient Tunnels (TGT)

Estimation of total N% of wheat grainsand straw

Chlorophyllcontent

Root length (cm) and Root weight (mg)

Coleoptile length(cm)

Stomata / cm2

Leaf area index (LAI)

Physiological parameters

Days to physiological maturity

T cell phenotypic distribution in HBsAgPositive, HBsAgNegative from HBsAg positive mothers and healthy newborns.

GFP-β-catenin T41A mutant

Reagents

Standard curve of ascorbic acid

ZnSO4

Procedur

Procedur

Procedure

Procedure

Reagent

Total antioxidant activit

Procedur

Material

Total dissolved solids

High Performance Liquid Chromatography (HPLC) analysis

High Performance Thin Layer Chromatography (HPTLC) analysis

. Thin Layer Chromatography (TLC) analysis

Preliminary phytochemical screening (Ali, 1998; Evans, 2002)

Microbial Contamina

Foaming Index

pH values

Ash values

Extractive value

. Physico-chemical standardization

Standardization of ethanolic extract of E.ribes (WHO, 1998; IndianPharmacopoeia, 1996)

Estimation of Stevioside

Estimation of Steviol

Estimation of total phenols

Mean Somatotypes:

Somatocharts:

Balanced mesomorph:

Purity of the DNA extracted from various environmental samples was confirmed by subjecting the extracted DNA to restriction digestion. DNA was digested with Sau3AI (New England Biolabs). One μg of metagenomic DNA in 20 μL reaction mixture was treated with 0.5 U of Sau3AI and incubated at 37 °Cfor 10 min. The reaction was terminated at 80 °C for 20 min and the digested DNA was fractionated on 1.2 % (w/v) agarose gel.

Restriction digestion

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.

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

Comparison of yield and purity of crude DNA

Various strains of Escherchia coli (DH5α, XL1Blue, DH10B) were used as hosts for the propagation of recombinant vectors. In addition, Bacillus subtilis was used as a host for the expression of xylanase gene from the recombinant vector pWHMxyl. Different vectors used in this investigation are listed in

BACTERIAL STRAINS

All the peptides used in this study were synthesized by solid phase method on an automated peptide synthesizer (Applied Biosystems, Model 431A), using F-moc (9-fluorenylmethyloxycarbonyl) chemistry on a p-hydroxymethyl phenoxymethyl polystyrene resin (Nova Biochem). For the peptide synthesis, 0.1 mmol of the resin was used and deprotected using 20% piperidine in N-methyl-pyrrolidone (NMP). Subsequently 0.5nmol of the first amino acid was added and coupling was performed usmg DCC-HoBt (dicyclohexylcarbodiimide-hydroxybezotriazole) ester formation method. All other amino acids were coupled by DCC ester coupling. Amino acids and solutions required for peptide synthesis were procured from Nova Biochem and Applied Biosystems, respectively. After completion of synthesis, deprotection was carried out in 20% piperidine/DMF. Finally, the resin was shrunk using ether and dried under vacuum for a minimum of four hours. The cleavage was performed in dark using 94% TF A, 5% anisole, EDT and water accompanied by continuous stirring for two hours. The resin was then filtered and washed with DCM and the solution was evaporated on a rotary evaporator (Buchi, Switzerland) till only a small quantity of DCM/cleavage mixture is left. Cold anhydrous diethyl ether was added to the filtrate to aid in the separation of scavengers from the mixture. The peptides were then extracted with water using a separating funnel. Extraction was followed by evaporation of residual diethyl ether on the rotary evaporator. Total aqueous layer was then frozen as a thin film and lyophilized.

Procedure for peptide synthesis

vector, under the phage T7 promoter, in BL21 (DE3) cells, and under the T5 phage promoter, in the pQE30 vector for expression in SG13009[pREP4] and M15[pREP4] cell strains. For cloning in pRSET B, the full length bZP3 initially subcloned in the pBacPAK8 vector at the Kpn I and Sac I sites was released after digestion with Kpn I and EcoR I and cloned in a similarly restricted pRSETB vector inframe with an N-terminal His6 tag. For cloning in the pQE30 vector, the pBacPAK8 carrying the full length bZP3 was initially digested with Not I, filled in with Klenow and then digested with Kpn I. The purified bZP3 fragment was then cloned in the vector digested with Kpn I and Sma I in frame with an N-terminal His6 tag. Though transformants positive for the bZP3 insert in the right reading frame were recovered, no expression could be detected by SDS-PAGE or immunoblots in either case. An alternate strategy was then devised in which an internal fragment of the gene, excluding the signal sequence and the transmembrane-like domain, following the putative furin cleavage site, was amplified by PCR using the forward primer 5'-CGGGATCCCAACCCTTCTGGCTCTTG-3' incorporating a BamH I site and the reverse primer 5'-CCGAGCTCAGAAGCAGACCTGGACCA-3' incorporating a Sac I site. The PCR was done in a 50 J!l volume using 50 pM of each primer and Vent polymerase for extension. The pBluescript-bZP3 (1 0 ng) having a full length bZP3 insert was used as the template and was initially denatured at 95°C for 10 min. Amplification was carried out for 35 cycles of denaturation at 95°C for 2 min, primer annealing at 600C for 2 min and extension at 72°C for 3 min followed by a final extension at 72oc .for 15 min. The amplified bZP3 fragment was digested with BamH I and Sac I and cloned in frame downstream of a His6 tag under the T5 promoter-lac operator control in the pQE30 vector. The authenticity of the construct was confirmed by N-terminal sequencing using an upstream sequencing primer GGCGT ATCACGAGGCCCTTTCG.

Our initial attempts to express the full length gene in E. coli as a His6 fusion protein failed. Attempts were initially made to express the His6-bZP3 protein in the pRSET B

PCR Amplification and Cloning in pQE30 Vector

ligation reactions were carried out usmg conditions and buffers specified by the manufacturer.

The PCR amplified eDNA fragment corresponding to bZP3 was resolved on a 0.8% agarose gel run using IX TAE buffer (0.04 M Tris-acetate, O.OOI M EDTA) and purified using the Geneclean® II kit. The PCR amplified bZP3 was digested with Kpn I and Sac I and ligated into the pBluescriptll SK(+) vector at the same sites. The digestion and

Agarose Gel Electrophoresis, Digestion and Ligation

PBS and then replenished with the complete medium. Two days following transfection, the cells were subcultured into the appropriate selective medium for selection of stable clones as described below.

Calcium phosphate mediated stable transfections were performed by the method of Graham and Van der Eb ( 1973 with modifications as described by Gorman ( 1986 ). For each plasmid, two petri dishes each containing 0. 5 x 106 CHO-K1 cells were used, with 10 ug of cesium purified DNA for each transfection. A mock transfection which did not contain any DNA, was performed simultaneously as negative control. Precipitation of the DNA was done with great care to ensure the obtention of a fine, translucent precipitate rather than a dense and opaque precipitate. The calcium phosphate I DNA precipitate was added in 4 ml medium to the cells and the cells incubated for 3 hours at 37°C. At this stage, the cells were examined under the microscope and a fine precipitate appeared as small grains all over the cells. The cells were washed once with serum free medium and a glycerol shock given for 3 minutes at 37°C. The cells were washed twice again with

Using calcium phosphate.

bands seen in the DNA size marker, were marked with a ball -point pen at the places where small holes had been pierced in the gel earlier ( see above ). Thus it was easy to monitor the size of the fragments showing hybridisation to the probe. The gel was then peeled off and the membrane w~shed in 6 X sse with gentle rocking for 10 minutes to wash away any residual agarose sticking to the membrane. After air drying at room temperature, the membrane was baked at so0e for two hours. The baked filter was stored at room temperature in a dessicator, if not used immediately. The dehydrated gel was restained in water containing 0.5 ug I ml ethidium bromide for 30 minutes and examined on a short wave UV transilluminator to check for the presence of any DNA fragments that escaped blotting. The absence of any residual bands indicated that the transfer was complete.

Restriction fragments of DNA resolved on agarose gel were transferred to nylon membrane ( GeneScreen or GeneScreen Plus by the capillary blotting procedure of Southern ( 1975 ) as described by Maniatis et al., ( 1982 ) . After the completion of electrophoresis, the gel was stained and photographed as described earlier. Position of the various bands obtained in the DNA size marker lane were marked by piercing small holes at the two ends of each band in the gel with a yellow tip. The gel was then denatured, neutralised and blotted essentially as described by Maniatis et al., ( 1982 ) . Locally available coarse absorbent paper was used to make the paper towels of the appropriate size. In case of genomic DNA from mammalian cells, the agarose gel was first treated with 0.25 M HCl for 10 minutes, followed by the rest of the procedure as mentioned above. The transfer buffer was 20 X SSPE in all cases. To prevent the absorption of fluid from the 3 MM paper under the gel directly to the blotting paper atop the nylon membrane, the gel was surrounded with polythene sheets to minimise the direct contact between the blotting paper and the 3 MM paper placed under the gel. The blotting was performed for 18 -24 hours. After the transfer was over, the paper towels and the 3 MM papers on top of the nylon filter were peeled off. The gel along with the attached membrane, was turned over and kept on a clean sheet of 3 MM paper with the gel side up. The position of the gel slots was marked with a ball -point pen. Also, the positions of the

southern blot.

lectrophoresed on 0.7 % -1.2 % agarose gels in TAE or TBE buffer. Choice of the percentage of agarose and the electrophoresis buffer system was made following the guidelines of Maniatis et al., ( 1982 ). In general, upto 1 kb fragments were resolved on 1.2 % agarose gels using TBE buffer. For most other purposes, TAE buffer was used. Agarose gel electrophoresis was carried out as described by Maniatis et al., ( 1982 ) . The run was stopped when the bromophenol blue dye migrated to within 1 em -1.5 em from the edge of ' the gel, except when the sample had fragments smaller than 500 bp, in which case the elctrophoresis was terminated at an earlier stage. The gel was immersed in water containing 0.5 ug I ml ethidium bromide, for 30 minutes, to stain the DNA. When detecting very low amounts of DNA, the staining was done for 60 minutes followed by destaining in 1 mM Mgso4 for one hour at room temperature. The DNA bands were visualised on a short wavelength UV transilluminator ( Fotodyne, Inc., USA and photographed with a Polaroid MP-4 camera using Polaroid type 667 film.

DNA digested with restriction enzymes was

ecanted and the pellet dried briefly under vacuum. The final DNA pellet was resuspended in 500 ul of TE. A 1:50 dilution of the sample was used to measure the absorbance at 260 nm and at 280 nm. The A260 and A280 values were used to estimate the concentration and purity of the sample as described by Maniatis et al., ( 1982).

further purified by centrifugation to equilibrium in a 30 ml cesium chloride -ethidium bromide density gradient, as described by Maniatis et al., ( 1982 ) . The band corresponding to closed circular plasmid DNA was collected and further purified by a second centrifugation to equilibrium in a 6. 5 ml cesium chloride -ethidium bromide density gradient. The final DNA band collected from the gradient was extracted with an equal volume of isopropanol which had been previously saturated with TE and cesium chloride. This extraction was repeated twice to completely remove the ethidium bromide from the DNA sample. The DNA was then dialysed against one liter of TE for at least 8 hours, at 4 °c, with several changes of TE. To the dialysed sample, one tenth volume of 3 M sodium acetate, pH 5.2, was added and the DNA precipitated with two volumes of chilled ethanol. The precipitation was carried out 0/N at 0 -20 c. The precipitated centrifugation at 10, 000 rpm, DNA was collected by for 10 minutes. The supernate was carefully d

resuspended in 20 ml of Tris -Glucose solution ( 25 mM Tris. HCl, pH 8. 0; 50 mM Glucose ) . The cells were vortexed followed by repeated pipetting to obtain a uniform cell suspension. To this, 6.0 ml of a freshly prepared lysozyme solution ( 10 mg 1 ml, prepared freshly in sterile distilled water ) was added. The cell suspension was swirled to mix thoroughly and incubated for 5 minutes at room temperature. Next, 0.5 M EDTA was added to a final concentration of 10 mM, the contents swirled to mix and incubated in ice for 20 minutes. Next, 40 ml of a lytic mix containing 0. 1 % SDS and 0. 2 N NaOH was added. This was prepared freshly by mixing 4 ml of 10 % SDS solution into 36 ml of 0.22 N NaOH solution. The solution was mixed by vigorous but brief shaking till the cell lysate became clear, followed by incubation on ice for 5 minutes. Finally, 20 ml of 5 M potassium acetate solution, pH 4.8 was added. Again the contents were swirled to mix, followed by incubation in ice for at least 1 - 2 hours. The lysate was centrifuged at 10,000 rpm for 30 minutes at 4°c. The supernate was filtered through sterilised glass wool kept in a funnel, and collected in a graduated cylinder. The measured volume of the cell lysate was transferred into another centrifuge bottle and two volumes of 95 % ethanol added to precipitate the DNA, at 0 -20 c, 0/N. The DNA was pelleted by centrifugation at 10,000 rpm at 4 °c for 30 minutes. The supernate was carefully poured off and the pellet res~spended in 25 ml of TE ( 10 mM Tris.HCl, pH 8.0; 1 mM EDTA ). The plasmid DNA was