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Glycine

Glycine is an important amino acid that plays a crucial role in various biological processes.
It is the smallest and simplest of the 20 standard amino acids, with a single hydrogen atom as its side chain.
Glycine is involved in the synthesis of proteins, the metabolism of fats and carbohydrates, and the functioning of the nervous system.
It is also a neurotransmitter in the central nervous system, where it inhibits neuronal excitation.
Glycine has been studied extensively for its potential therapeutic applications, including in the treatment of neurological disorders, metabolic conditions, and immune-related diseases.
The PubCompare.ai platform can help researchers optimize their Glycine studies by providing access to a wealth of protocols from literature, preprints, and patents, and leveraging AI-driven comparisons to identify the best protocols and products.
This can enhance reproducibility and accuracy in Glycine research, leading to more reliable and impactful findings.

Most cited protocols related to «Glycine»

Whole-genome protein sequences from Arabidopsis, Populus, Vitis, Glycine, Oryza, Brachypodium, Sorghum and Zea were merged and searched against themselves for homology using BLASTP with an E-value cutoff of 10−5. Default parameters of OrthoMCL (50 (link)) were used. The combination of OrthoMCL intermediate files ‘orthologs.txt’ and ‘coorthologs.txt’ (generated by orthomclDumpPairsFiles) was used as the whole set of ortholog pairs.
Publication 2012
Amino Acid Sequence Arabidopsis Brachypodium Genome Glycine Oryza Populus Sorghum Vitis
Quantitation limit was tested by analysis of six consecutive injections of salicylic acid standard solution at the concentration of 0.0005 mg ml−1 in a presence of placebo and glycine. Mean, SD and %RSD of the peak area response were calculated.
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Publication 2022
Glycine Placebos Salicylic Acid
Acetyl and N-methyl capped dipeptides of the natural amino acids, except proline, alanine, and glycine, were built using LEaP29 at α (−60°, −45°) and β (−135°, 135°) backbone conformations.
χ was explored by rotating in 10° increments, re-optimizing at each step, or by high temperature simulation (described in Results).
Quantum mechanics optimizations were performed with RHF/6-31G*. Scanned residues were optimized using GAMESS (US)30 with default options. Optimization continued until the RMS gradient was less than 1.0 × 10−4 Hartree Bohr−1, with an initial trust radius of 0.1 Bohr that could then adjust between 0.05 and 0.5 Bohr. Minimization proceeded by the quadratic approximation. Residues sampled by high temperature simulations were optimized using Gaussian9831 with VTight convergence criteria. Quantum mechanics energies for training data were calculated with MP2/6-31+G**. Molecular mechanics re-optimizations were performed in the gas phase with ff99SB for a maximum of 1.0 × 107 (link) cycles or until the RMS gradient was less than 1.0 × 10−4 kcal mol−1 Å−1, with a non-bonded cutoff of 99.0 Å and initial step size of 10−4. Dihedral restraint force constants were 2.0 × 105 kcal mol−1 rad−2. Minimization employed 10 steps of steepest descent followed by conjugate gradient. Molecular mechanics energies were calculated from the last step of ff99SB minimization.
Publication 2015
Alanine Amino Acids Dipeptides Fever Glycine Mechanics Proline Radius STEEP1 protein, human Vertebral Column
One hour after infecting the cell monolayers with 30–50 plaque forming units of the virus in 1 ml of maintenance medium without trypsin, we removed the virus inoculum, covered the cells with 3 ml of the different overlay media and incubated cultures at 35°C in 5% CO2 atmosphere. In the case of MC and Avicel overlays, care was taken not to disturb the plates during the incubation period in order to avoid formation of non-even plaques. After three days of incubation, we removed the overlays and fixed the cells. Agar overlay was removed using metal spatula; MC, Avicel, and liquid overlays were removed by suction. The cells were fixed with 4% paraformaldehyde solution in MEM for 30 min at 4°C and washed with PBS. All subsequent treatments of the cells were performed at room temperature. We permeabilized the cells and simultaneously blocked residual aldehyde groups by incubating the cells for 10–20 min with 1 ml/well of solution containing 0.5 % Triton-X-100 and 20 mM glycine in PBS. We immuno-stained virus-infected cells by incubating for 1 hr with monoclonal antibodies specific for the influenza A virus nucleoprotein (kindly provided by Dr. Alexander Klimov at Centers for Disease Control, USA) followed by 1 hr incubation with peroxidase-labeled anti-mouse antibodies (DAKO, Denmark) and 30 min incubation with precipitate-forming peroxidase substrates. Solution of 10% normal horse serum and 0.05% Tween-80 in PBS was used for the preparation of working dilutions of immuno-reagents. We washed the cells after the primary and secondary antibodies by incubating them three times for 3–5 min with 0.05% Tween-80 in PBS. As peroxidase substrates, we employed either ready to use True Blue™ (KPL) or solution of aminoethylcarbazole (AEC, Sigma) (0.4 mg/ml) prepared in 0.05 M sodium acetate buffer, pH 5.5 and containing 0.03% H2O2. Stained plates were washed with tap water to stop the reaction and dried. In the case of True Blue staining, which is relatively unstable in water solutions, plates were dried inverted in order to minimize bleaching. Stained plates were scanned on a flat bed scanner and the data were acquired by Adobe Photoshop 7.0 software.
As an alternative to immuno-staining, in some experiments we revealed plaques as areas of destroyed cells. To this end, after removing the overlays, we stained the cells with 1% crystal violet solution in 20% methanol in water.
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Publication 2006
Agar Aldehydes Anti-Antibodies Antibodies Atmosphere Avicel Buffers Dental Plaque Equus caballus Glycine Metals Methanol Monoclonal Antibodies Mus NP protein, Influenza A virus paraform Peroxidase Peroxide, Hydrogen Senile Plaques Serum Sodium Acetate Suction Drainage Technique, Dilution Triton X-100 true blue Trypsin Tween 80 Violet, Gentian Virus
We identified eight publicly available MHC class II prediction tools through literature search and the IMGT link list at http://imgt.cines.fr/textes/IMGTbloc-notes/. For each tool, we mapped the MHC types for which predictions could be made to the four-digit HLA nomenclature (e.g., HLA-DRB1*0101). If this mapping could not be done exactly, we left that type/tool combination out of the evaluation. For example, HLA-DR4 could refer to HLA-DRB1*0401, DRB1*0402 etc, which do have distinct binding specificities.
For the ARB evaluation, the 10-fold cross validation results stored at IEDB was used to estimate performance since ARB was trained on datasets overlapping with the one used in this study. For the other seven tools in the evaluation, we wrote python script wrappers to automate prediction retrieval. For the SYFPEITHI prediction, we patched each testing peptide with three Glycine residues at both ends before we submitted it for prediction. This was recommended by the creators of SYFPEITHI method to ensure that all potential binders are presented to the prediction algorithm. For all other methods, the original testing peptides were submitted directly for prediction. Peptide sequences were sent to the web servers one at a time and predictions were extracted from the server's response. To assign a single prediction for peptides longer than nine amino acids in the context of tools predicting the affinity of 9-mer core binding regions, we took the highest affinity prediction of all possible 9-mers within the longer peptide as the prediction result.
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Publication 2008
Amino Acids Fingers Genes, MHC Class II Glycine GPER protein, human HLA-DR4 Antigen HLA-DRB1 Antigen Peptides Python

Most recents protocols related to «Glycine»

Example 8

Characterization of Absorption, Distribution, Metabolism, and Excretion of Oral [14C]Vorasidenib with Concomitant Intravenous Microdose Administration of [13C315N3]Vorasidenib in Humans

Metabolite profiling and identification of vorasidenib (AG-881) was performed in plasma, urine, and fecal samples collected from five healthy subjects after a single 50-mg (100 μCi) oral dose of [14C]AG-881 and concomitant intravenous microdose of [13C3 15N3]AG-881.

Plasma samples collected at selected time points from 0 through 336 hour postdose were pooled across subjects to generate 0—to 72 and 96-336-hour area under the concentration-time curve (AUC)-representative samples. Urine and feces samples were pooled by subject to generate individual urine and fecal pools. Plasma, urine, and feces samples were extracted, as appropriate, the extracts were profiled using high performance liquid chromatography (HPLC), and metabolites were identified by liquid chromatography-mass spectrometry (LC-MS and/or LC-MS/MS) analysis and by comparison of retention time with reference standards, when available.

Due to low radioactivity in samples, plasma metabolite profiling was performed by using accelerator mass spectrometry (AMS). In plasma, AG-881 was accounted for 66.24 and 29.47% of the total radioactivity in the pooled AUC0-72 h and AUC96-336 h plasma, respectively. The most abundant radioactive peak (P7; M458) represented 0.10 and 43.92% of total radioactivity for pooled AUC0-72 and AUC96-336 h plasma, respectively. All other radioactive peaks accounted for less than 6% of the total plasma radioactivity and were not identified.

The majority of the radioactivity recovered in feces was associated with unchanged AG-881 (55.5% of the dose), while no AG-881 was detected in urine. In comparison, metabolites in excreta accounted for approximately 18% of dose in feces and for approximately 4% of dose in urine. M515, M460-1, M499, M516/M460-2, and M472/M476 were the most abundant metabolites in feces, and each accounted for approximately 2 to 5% of the radioactive dose, while M266 was the most abundant metabolite identified in urine and accounted for a mean of 2.54% of the dose. The remaining radioactive components in urine and feces each accounted for <1% of the dose.

Overall, the data presented indicate [14C]AG-881 underwent moderate metabolism after a single oral dose of 50-mg (100 μCi) and was eliminated in humans via a combination of metabolism and excretion of unchanged parent. AG-881 metabolism involved the oxidation and conjugation with glutathione (GSH) by displacement of the chlorine at the chloropyridine moiety. Subsequent biotransformation of GSH intermediates resulted in elimination of both glutamic acid and glycine to form the cysteinyl conjugates (M515 and M499). The cysteinyl conjugates were further converted by a series of biotransformation reactions such as oxidation, S-dealkylation, S-methylation, S-oxidation, S-acetylation and N-dealkylation resulting in the formation multiple metabolites.

A summary of the metabolites observed is included in Table 2

TABLE 2
Retention
ComponentTimeMatrix
designation(Minutes)[M + H]+Type of BiotransformationPlasmaUrineFeces
Unidentified 17.00UnknownX
M2667.67a267N-dealkylationX
Unidentified 2UnknownX
Unidentified 3UnknownX
Unidentified 4UnknownX
Unidentified 5UnknownX
M51519.79b516OxidationX
M460-120.76b461OxidationX
M49921.22b500Dechloro-glutathioneXX
conjugation + hydrolysis
M51621.89b517Oxidative-deaminationX
M460-221.98b461OxidationX
M47222.76b473S-dealkylation + S-X
acetylation + reduction
M47622.76b477OxidationX
Unidentified 6UnknownX
M47423.63b475OxidationX
Unidentified 7UnknownX
M43025.88b431AG-881-oxidationX
M42630.62b427S-dealkylation + methylationX
M45831.03c459AG-69460X*
AG-88139.41b415AG-881XX
M42847.40b429S-dealkylation + oxidationX
Table 3 contains a summary of protonated molecular ions and characteristic product ions for AG-881 and identified metabolites

TABLE 3
RetentionCharacteristic
MetaboliteTimeProposed MetaboliteProduct Ions
designation(Minutes)[M + H]+Identification(m/z)Matrix
M266 7.88a267[Figure (not displayed)]
188, 187Urine
M51519.79b516[Figure (not displayed)]
429, 260, 164, 153Feces
M460-120.76b461[Figure (not displayed)]
379, 260, 164Feces
M49921.22b500[Figure (not displayed)]
437, 413, 260, 164, 137Urine Feces
M51621.89b517[Figure (not displayed)]
427, 260, 164, 153Feces
M460-221.98b461[Figure (not displayed)]
369, 260, 164, 139, 121, 93Feces
M47222.76b473[Figure (not displayed)]
429, 260, 179, 164, 153Feces
M47622.76b477[Figure (not displayed)]
395, 260, 164, 139, 119Feces
M47423.63b475[Figure (not displayed)]
260, 164, 68Feces
M43025.88b431[Figure (not displayed)]
260, 164, 155, 68Feces
M42630.62b427[Figure (not displayed)]
260, 164, 151Feces
M45831.03b459[Figure (not displayed)]
380, 311, 260, 183, 164, 130Plasma Fecesd
AG-88139.41b415[Figure (not displayed)]
319, 277, 260, 240, 164, 139, 119, 68Plasma Fecesd
M42847.40b429[Figure (not displayed)]
260, 164, 153Feces
Notes
aRetention time from analysis of a urine sample
bRetention time from analysis of a feces sample
cRetention time from analysis of a plasma sample
dM458 was only detected in feces by mass spectrometry, not by radioprofiling.
The proposed (theoretical) biotransformation pathways leading to the observed metabolites are shown in FIG. 1.

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Patent 2024
Acetylation AG 30 Biotransformation Chlorine Dealkylation Deamination Elements, Radioactive Feces Glutamic Acid Glutathione Glycine Healthy Volunteers High-Performance Liquid Chromatographies Homo sapiens Hydrolysis Intravenous Infusion Ions Liquid Chromatography Mass Spectrometry Metabolism Methylation Parent Plasma Radioactivity Retention (Psychology) Tandem Mass Spectrometry Urinalysis Urine vorasidenib

Example 2

A. Seed Treatment with Isolated Microbe

In this example, an isolated microbe from Tables 1-3 will be applied as a seed coating to seeds of corn (Zea mays). Upon applying the isolated microbe as a seed coating, the corn will be planted and cultivated in the standard manner.

A control plot of corn seeds, which did not have the isolated microbe applied as a seed coating, will also be planted.

It is expected that the corn plants grown from the seeds treated with the seed coating will exhibit a quantifiably higher biomass than the control corn plants.

The biomass from the treated plants may be about 1-10% higher, 10-20% higher, 20-30% higher, 30-40% higher, 40-50% higher, 50-60% higher, 60-70% higher, 70-80% higher, 80-90% higher, or more.

The biomass from the treated plants may equate to about a 1 bushel per acre increase over the controls, or a 2 bushel per acre increase, or a 3 bushel per acre increase, or a 4 bushel per acre increase, or a 5 bushel per acre increase, or more.

In some aspects, the biomass increase is statistically significant. In other aspects, the biomass increase is not statistically significant, but is still quantifiable.

B. Seed Treatment with Microbial Consortia

In this example, a microbial consortium, comprising at least two microbes from Tables 1-3 will be applied as a seed coating to seeds of corn (Zea mays). Upon applying the microbial consortium as a seed coating, the corn will be planted and cultivated in the standard manner.

A control plot of corn seeds, which did not have the microbial consortium applied as a seed coating, will also be planted.

It is expected that the corn plants grown from the seeds treated with the seed coating will exhibit a quantifiably higher biomass than the control corn plants.

The biomass from the treated plants may be about 1-10% higher, 10-20% higher, 20-30% higher, 30-40% higher, 40-50% higher, 50-60% higher, 60-70% higher, 70-80% higher, 80-90% higher, or more.

The biomass from the treated plants may equate to about a 1 bushel per acre increase over the controls, or a 2 bushel per acre increase, or a 3 bushel per acre increase, or a 4 bushel per acre increase, or a 5 bushel per acre increase, or more.

In some aspects, the biomass increase is statistically significant. In other aspects, the biomass increase is not statistically significant, but is still quantifiable.

C. Treatment with Agricultural Composition Comprising Isolated Microbe

In this example, an isolated microbe from Tables 1-3 will be applied as an agricultural composition, administered to the corn seed at the time of sowing.

For example, it is anticipated that a farmer will apply the agricultural composition to the corn seeds simultaneously upon planting the seeds into the field. This can be accomplished, for example, by applying the agricultural composition to a hopper/bulk tank on a standard 16 row planter, which contains the corn seeds and which is configured to plant the same into rows. Alternatively, the agricultural composition can be contained in a separate bulk tank on the planter and sprayed into the rows upon planting the corn seed.

A control plot of corn seeds, which are not administered the agricultural composition, will also be planted.

It is expected that the corn plants grown from the seeds treated with the agricultural composition will exhibit a quantifiably higher biomass than the control corn plants.

The biomass from the treated plants may be about 1-10% higher, 10-20% higher, 20-30% higher, 30-40% higher, 40-50% higher, 50-60% higher, 60-70% higher, 70-80% higher, 80-90% higher, or more.

The biomass from the treated plants may equate to about a 1 bushel per acre increase over the controls, or a 2 bushel per acre increase, or a 3 bushel per acre increase, or a 4 bushel per acre increase, or a 5 bushel per acre increase, or more.

In some aspects, the biomass increase is statistically significant. In other aspects, the biomass increase is not statistically significant, but is still quantifiable.

D. Treatment with Agricultural Composition Comprising Microbial Consortia

In this example, a microbial consortium, comprising at least two microbes from Tables 1-3 will be applied as an agricultural composition, administered to the corn seed at the time of sowing.

For example, it is anticipated that a farmer will apply the agricultural composition to the corn seeds simultaneously upon planting the seeds into the field. This can be accomplished, for example, by applying the agricultural composition to a hopper/bulk tank on a standard 16 row planter, which contains the corn seeds and which is configured to plant the same into rows. Alternatively, the agricultural composition can be contained in a separate bulk tank on the planter and sprayed into the rows upon planting the corn seed.

A control plot of corn seeds, which are not administered the agricultural composition, will also be planted.

It is expected that the corn plants grown from the seeds treated with the agricultural composition will exhibit a quantifiably higher biomass than the control corn plants.

The biomass from the treated plants may be about 1-10% higher, 10-20% higher, 20-30% higher, 30-40% higher, 40-50% higher, 50-60% higher, 60-70% higher, 70-80% higher, 80-90% higher, or more.

The biomass from the treated plants may equate to about a 1 bushel per acre increase over the controls, or a 2 bushel per acre increase, or a 3 bushel per acre increase, or a 4 bushel per acre increase, or a 5 bushel per acre increase, or more.

In some aspects, the biomass increase is statistically significant. In other aspects, the biomass increase is not statistically significant, but is still quantifiable.

A. Seed Treatment with Isolated Microbe

In this example, an isolated microbe from Tables 1-3 will be applied as a seed coating to seeds of corn (Zea mays). Upon applying the isolated microbe as a seed coating, the corn will be planted and cultivated in the standard manner.

A control plot of corn seeds, which did not have the isolated microbe applied as a seed coating, will also be planted.

It is expected that the corn plants grown from the seeds treated with the seed coating will exhibit a quantifiable and superior ability to tolerate drought conditions and/or exhibit superior water use efficiency, as compared to the control corn plants.

The drought tolerance and/or water use efficiency can be based on any number of standard tests from the art, e.g leaf water retention, turgor loss point, rate of photosynthesis, leaf color and other phenotypic indications of drought stress, yield performance, and various root morphological and growth patterns.

B. Seed Treatment with Microbial Consortia

In this example, a microbial consortium, comprising at least two microbes from Tables 1-3 will be applied as a seed coating to seeds of corn (Zea mays). Upon applying the microbial consortium as a seed coating, the corn will be planted and cultivated in the standard manner.

A control plot of corn seeds, which did not have the microbial consortium applied as a seed coating, will also be planted.

It is expected that the corn plants grown from the seeds treated with the seed coating will exhibit a quantifiable and superior ability to tolerate drought conditions and/or exhibit superior water use efficiency, as compared to the control corn plants.

The drought tolerance and/or water use efficiency can be based on any number of standard tests from the art, e.g leaf water retention, turgor loss point, rate of photosynthesis, leaf color and other phenotypic indications of drought stress, yield performance, and various root morphological and growth patterns.

C. Treatment with Agricultural Composition Comprising Isolated Microbe

In this example, an isolated microbe from Tables 1-3 will be applied as an agricultural composition, administered to the corn seed at the time of sowing.

For example, it is anticipated that a farmer will apply the agricultural composition to the corn seeds simultaneously upon planting the seeds into the field. This can be accomplished, for example, by applying the agricultural composition to a hopper/bulk tank on a standard 16 row planter, which contains the corn seeds and which is configured to plant the same into rows. Alternatively, the agricultural composition can be contained in a separate bulk tank on the planter and sprayed into the rows upon planting the corn seed.

A control plot of corn seeds, which are not administered the agricultural composition, will also be planted.

It is expected that the corn plants grown from the seeds treated with the with the agricultural composition will exhibit a quantifiable and superior ability to tolerate drought conditions and/or exhibit superior water use efficiency, as compared to the control corn plants.

The drought tolerance and/or water use efficiency can be based on any number of standard tests from the art, e.g leaf water retention, turgor loss point, rate of photosynthesis, leaf color and other phenotypic indications of drought stress, yield performance, and various root morphological and growth patterns.

D. Treatment with Agricultural Composition Comprising Microbial Consortia

In this example, a microbial consortium, comprising at least two microbes from Tables 1-3 will be applied as an agricultural composition, administered to the corn seed at the time of sowing.

For example, it is anticipated that a farmer will apply the agricultural composition to the corn seeds simultaneously upon planting the seeds into the field. This can be accomplished, for example, by applying the agricultural composition to a hopper/bulk tank on a standard 16 row planter, which contains the corn seeds and which is configured to plant the same into rows. Alternatively, the agricultural composition can be contained in a separate bulk tank on the planter and sprayed into the rows upon planting the corn seed.

A control plot of corn seeds, which are not administered the agricultural composition, will also be planted.

It is expected that the corn plants grown from the seeds treated with the with the agricultural composition will exhibit a quantifiable and superior ability to tolerate drought conditions and/or exhibit superior water use efficiency, as compared to the control corn plants.

The drought tolerance and/or water use efficiency can be based on any number of standard tests from the art, e.g leaf water retention, turgor loss point, rate of photosynthesis, leaf color and other phenotypic indications of drought stress, yield performance, and various root morphological and growth patterns.

A. Seed Treatment with Isolated Microbe

In this example, an isolated microbe from Tables 1-3 will be applied as a seed coating to seeds of corn (Zea mays). Upon applying the isolated microbe as a seed coating, the corn will be planted and cultivated in the standard manner.

A control plot of corn seeds, which did not have the isolated microbe applied as a seed coating, will also be planted.

It is expected that the corn plants grown from the seeds treated with the seed coating will exhibit a quantifiable and superior ability to utilize nitrogen, as compared to the control corn plants.

The nitrogen use efficiency can be quantified by recording a measurable change in any of the main nitrogen metabolic pool sizes in the assimilation pathways (e.g., a measurable change in one or more of the following: nitrate, nitrite, ammonia, glutamic acid, aspartic acid, glutamine, asparagine, lysine, leucine, threonine, methionine, glycine, tryptophan, tyrosine, total protein content of a plant part, total nitrogen content of a plant part, and/or chlorophyll content), or where the treated plant is shown to provide the same or elevated biomass or harvestable yield at lower nitrogen fertilization levels compared to the control plant, or where the treated plant is shown to provide elevated biomass or harvestable yields at the same nitrogen fertilization levels compared to a control plant.

B. Seed Treatment with Microbial Consortia

In this example, a microbial consortium, comprising at least two microbes from Tables 1-3 will be applied as a seed coating to seeds of corn (Zea mays). Upon applying the microbial consortium as a seed coating, the corn will be planted and cultivated in the standard manner.

A control plot of corn seeds, which did not have the microbial consortium applied as a seed coating, will also be planted.

It is expected that the corn plants grown from the seeds treated with the seed coating will exhibit a quantifiable and superior ability to utilize nitrogen, as compared to the control corn plants.

The nitrogen use efficiency can be quantified by recording a measurable change in any of the main nitrogen metabolic pool sizes in the assimilation pathways (e.g., a measurable change in one or more of the following: nitrate, nitrite, ammonia, glutamic acid, aspartic acid, glutamine, asparagine, lysine, leucine, threonine, methionine, glycine, tryptophan, tyrosine, total protein content of a plant part, total nitrogen content of a plant part, and/or chlorophyll content), or where the treated plant is shown to provide the same or elevated biomass or harvestable yield at lower nitrogen fertilization levels compared to the control plant, or where the treated plant is shown to provide elevated biomass or harvestable yields at the same nitrogen fertilization levels compared to a control plant.

C. Treatment with Agricultural Composition Comprising Isolated Microbe

In this example, an isolated microbe from Tables 1-3 will be applied as an agricultural composition, administered to the corn seed at the time of sowing.

For example, it is anticipated that a farmer will apply the agricultural composition to the corn seeds simultaneously upon planting the seeds into the field. This can be accomplished, for example, by applying the agricultural composition to a hopper/bulk tank on a standard 16 row planter, which contains the corn seeds and which is configured to plant the same into rows. Alternatively, the agricultural composition can be contained in a separate bulk tank on the planter and sprayed into the rows upon planting the corn seed.

A control plot of corn seeds, which are not administered the agricultural composition, will also be planted.

It is expected that the corn plants grown from the seeds treated with the agricultural composition will exhibit a quantifiable and superior ability to utilize nitrogen, as compared to the control corn plants.

The nitrogen use efficiency can be quantified by recording a measurable change in any of the main nitrogen metabolic pool sizes in the assimilation pathways (e.g., a measurable change in one or more of the following: nitrate, nitrite, ammonia, glutamic acid, aspartic acid, glutamine, asparagine, lysine, leucine, threonine, methionine, glycine, tryptophan, tyrosine, total protein content of a plant part, total nitrogen content of a plant part, and/or chlorophyll content), or where the treated plant is shown to provide the same or elevated biomass or harvestable yield at lower nitrogen fertilization levels compared to the control plant, or where the treated plant is shown to provide elevated biomass or harvestable yields at the same nitrogen fertilization levels compared to a control plant.

D. Treatment with Agricultural Composition Comprising Microbial Consortia

In this example, a microbial consortium, comprising at least two microbes from Tables 1-3 will be applied as an agricultural composition, administered to the corn seed at the time of sowing.

For example, it is anticipated that a farmer will apply the agricultural composition to the corn seeds simultaneously upon planting the seeds into the field. This can be accomplished, for example, by applying the agricultural composition to a hopper/bulk tank on a standard 16 row planter, which contains the corn seeds and which is configured to plant the same into rows. Alternatively, the agricultural composition can be contained in a separate bulk tank on the planter and sprayed into the rows upon planting the corn seed.

A control plot of corn seeds, which are not administered the agricultural composition, will also be planted.

It is expected that the corn plants grown from the seeds treated with the agricultural composition will exhibit a quantifiable and superior ability to utilize nitrogen, as compared to the control corn plants.

The nitrogen use efficiency can be quantified by recording a measurable change in any of the main nitrogen metabolic pool sizes in the assimilation pathways (e.g., a measurable change in one or more of the following: nitrate, nitrite, ammonia, glutamic acid, aspartic acid, glutamine, asparagine, lysine, leucine, threonine, methionine, glycine, tryptophan, tyrosine, total protein content of a plant part, total nitrogen content of a plant part, and/or chlorophyll content), or where the treated plant is shown to provide the same or elevated biomass or harvestable yield at lower nitrogen fertilization levels compared to the control plant, or where the treated plant is shown to provide elevated biomass or harvestable yields at the same nitrogen fertilization levels compared to a control plant.

The inoculants were prepared from isolates grown as spread plates on R2A incubated at 25° C. for 48 to 72 hours. Colonies were harvested by blending with sterile distilled water (SDW) which was then transferred into sterile containers. Serial dilutions of the harvested cells were plated and incubated at 25° C. for 24 hours to estimate the number of colony forming units (CFU) in each suspension. Dilutions were prepared using individual isolates or blends of isolates (consortia) to deliver 1×105 cfu/microbe/seed and seeds inoculated by either imbibition in the liquid suspension or by overtreatment with 5% vegetable gum and oil.

Seeds corresponding to the plants of table 15 were planted within 24 to 48 hours of treatment in agricultural soil, potting media or inert growing media. Plants were grown in small pots (28 mL to 200 mL) in either a controlled environment or in a greenhouse. Chamber photoperiod was set to 16 hours for all experiments on all species. Air temperature was typically maintained between 22-24° C.

Unless otherwise stated, all plants were watered with tap water 2 to 3 times weekly. Growth conditions were varied according to the trait of interest and included manipulation of applied fertilizer, watering regime and salt stress as follows:

    • Low N—seeds planted in soil potting media or inert growing media with no applied N fertilizer
    • Moderate N—seeds planted in soil or growing media supplemented with commercial N fertilizer to equivalent of 135 kg/ha applied N
    • Insol P—seeds planted in potting media or inert growth substrate and watered with quarter strength Pikovskaya's liquid medium containing tri-calcium phosphate as the only form phosphate fertilizer.
    • Cold Stress—seeds planted in soil, potting media or inert growing media and incubated at 10° C. for one week before being transferred to the plant growth room.
    • Salt stress—seeds planted in soil, potting media or inert growing media and watered with a solution containing between 100 to 200 mg/L NaCl.

Untreated (no applied microbe) controls were prepared for each experiment. Plants were randomized on trays throughout the growth environment. Between 10 and 30 replicate plants were prepared for each treatment in each experiment. Phenotypes were measured during early vegetative growth, typically before the V3 developmental stage and between 3 and 6 weeks after sowing. Foliage was cut and weighed. Roots were washed, blotted dry and weighed. Results indicate performance of treatments against the untreated control.

TABLE 15
StrainShootRoot
Microbe sp.IDCropAssayIOC (%)IOC (%)
Bosea thiooxidans123EfficacyEfficacy
overall100%100%
Bosea thiooxidans54522WheatEarly vigor - insol P30-40 
Bosea thiooxidans54522RyegrassEarly vigor50-60 50-60 
Bosea thiooxidans54522RyegrassEarly vigor - moderate P0-100-10
Duganella violaceinigra111EfficacyEfficacy
overall100%100%
Duganella violaceinigra66361TomatoEarly vigor0-100-10
Duganella violaceinigra66361TomatoEarly vigor30-40 40-50 
Duganella violaceinigra66361TomatoEarly vigor20-30 20-30 
Herbaspirillum huttiense222Efficacy
overall100%
Herbaspirillum huttiense54487WheatEarly vigor - insol P30-40 
Herbaspirillum huttiense60507MaizeEarly vigor - salt stress0-100-10
Janthinobacterium sp.222Efficacy
Overall100%
Janthinobacterium sp.54456WheatEarly vigor - insol P30-40 
Janthinobacterium sp.54456WheatEarly vigor - insol P0-10
Janthinobacterium sp.63491RyegrassEarly vigor - drought0-100-10
stress
Massilia niastensis112EfficacyEfficacy
overall80%80%
Massilia niastensis55184WheatEarly vigor - salt stress0-1020-30 
Massilia niastensis55184WinterEarly vigor - cold stress0-1010-20 
wheat
Massilia niastensis55184WinterEarly vigor - cold stress20-30 20-30 
wheat
Massilia niastensis55184WinterEarly vigor - cold stress10-20 10-20 
wheat
Massilia niastensis55184WinterEarly vigor - cold stress<0<0
wheat
Novosphingobium rosa211EfficacyEfficacy
overall100%100%
Novosphingobium rosa65589MaizeEarly vigor - cold stress0-100-10
Novosphingobium rosa65619MaizeEarly vigor - cold stress0-100-10
Paenibacillus amylolyticus111EfficacyEfficacy
overall100%100%
Paenibacillus amylolyticus66316TomatoEarly vigor0-100-10
Paenibacillus amylolyticus66316TomatoEarly vigor10-20 10-20 
Paenibacillus amylolyticus66316TomatoEarly vigor0-100-10
Pantoea agglomerans323EfficacyEfficacy
33%50%
Pantoea agglomerans54499WheatEarly vigor - insol P40-50 
Pantoea agglomerans57547MaizeEarly vigor - low N<00-10
Pantoea vagans55529MaizeEarly vigor<0<0
(formerly P. agglomerans)
Polaromonas ginsengisoli111EfficacyEfficacy
66%100%
Polaromonas ginsengisoli66373TomatoEarly vigor0-100-10
Polaromonas ginsengisoli66373TomatoEarly vigor20-30 30-40 
Polaromonas ginsengisoli66373TomatoEarly vigor<010-20 
Pseudomonas fluorescens122Efficacy
100%
Pseudomonas fluorescens54480WheatEarly vigor - insol P>100 
Pseudomonas fluorescens56530MaizeEarly vigor - moderate N0-10
Rahnella aquatilis334EfficacyEfficacy
80%63%
Rahnella aquatilis56532MaizeEarly vigor - moderate N10-20 
Rahnella aquatilis56532MaizeEarly vigor - moderate N0-100-10
Rahnella aquatilis56532WheatEarly vigor - cold stress0-1010-20 
Rahnella aquatilis56532WheatEarly vigor - cold stress<00-10
Rahnella aquatilis56532WheatEarly vigor - cold stress10-20 <0
Rahnella aquatilis57157RyegrassEarly vigor<0
Rahnella aquatilis57157MaizeEarly vigor - low N0-100-10
Rahnella aquatilis57157MaizeEarly vigor - low N0-10<0
Rahnella aquatilis58013MaizeEarly vigor0-1010-20 
Rahnella aquatilis58013MaizeEarly vigor - low N0-10<0
Rhodococcus erythropolis313Efficacy
66%
Rhodococcus erythropolis54093MaizeEarly vigor - low N40-50 
Rhodococcus erythropolis54299MaizeEarly vigor - insol P>100 
Rhodococcus erythropolis54299MaizeEarly vigor<0<0
Stenotrophomonas chelatiphaga611EfficacyEfficacy
60%60%
Stenotrophomonas chelatiphaga54952MaizeEarly vigor0-100-10
Stenotrophomonas chelatiphaga47207MaizeEarly vigor<0 0
Stenotrophomonas chelatiphaga64212MaizeEarly vigor0-1010-20 
Stenotrophomonas chelatiphaga64208MaizeEarly vigor0-100-10
Stenotrophomonas chelatiphaga58264MaizeEarly vigor<0<0
Stenotrophomonas maltophilia612EfficacyEfficacy
43%66%
Stenotrophomonas maltophilia54073MaizeEarly vigor - low N50-60 
Stenotrophomonas maltophilia54073MaizeEarly vigor<00-10
Stenotrophomonas maltophilia56181MaizeEarly vigor0-10<0
Stenotrophomonas maltophilia54999MaizeEarly vigor0-100-10
Stenotrophomonas maltophilia54850MaizeEarly vigor 00-10
Stenotrophomonas maltophilia54841MaizeEarly vigor<00-10
Stenotrophomonas maltophilia46856MaizeEarly vigor<0<0
Stenotrophomonas rhizophila811EfficacyEfficacy
12.5%37.5%
Stenotrophomonas rhizophila50839MaizeEarly vigor<0<0
Stenotrophomonas rhizophila48183MaizeEarly vigor<0<0
Stenotrophomonas rhizophila45125MaizeEarly vigor<0<0
Stenotrophomonas rhizophila46120MaizeEarly vigor<00-10
Stenotrophomonas rhizophila46012MaizeEarly vigor<0<0
Stenotrophomonas rhizophila51718MaizeEarly vigor0-100-10
Stenotrophomonas rhizophila66478MaizeEarly vigor<0<0
Stenotrophomonas rhizophila65303MaizeEarly vigor<00-10
Stenotrophomonas terrae221EfficacyEfficacy
50%50%
Stenotrophomonas terrae68741MaizeEarly vigor<0<0
Stenotrophomonas terrae68599MaizeEarly vigor<00-10
Stenotrophomonas terrae68599Capsicum *Early vigor20-30 20-30 
Stenotrophomonas terrae68741Capsicum *Early vigor10-20 20-30 

The data presented in table 15 describes the efficacy with which a microbial species or strain can change a phenotype of interest relative to a control run in the same experiment. Phenotypes measured were shoot fresh weight and root fresh weight for plants growing either in the absence of presence of a stress (assay). For each microbe species, an overall efficacy score indicates the percentage of times a strain of that species increased a both shoot and root fresh weight in independent evaluations. For each species, the specifics of each independent assay is given, providing a strain ID (strain) and the crop species the assay was performed on (crop). For each independent assay the percentage increase in shoot and root fresh weight over the controls is given.

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Patent 2024
Ammonia Asparagine Aspartic Acid Biological Assay Bosea thiooxidans Calcium Phosphates Capsicum Cells Chlorophyll Cold Shock Stress Cold Temperature Crop, Avian Dietary Fiber DNA Replication Droughts Drought Tolerance Embryophyta Environment, Controlled Farmers Fertilization Glutamic Acid Glutamine Glycine Growth Disorders Herbaspirillum Herbaspirillum huttiense Leucine Lolium Lycopersicon esculentum Lysine Maize Massilia niastensis Methionine Microbial Consortia Nitrates Nitrites Nitrogen Novosphingobium rosa Paenibacillus Paenibacillus amylolyticus Pantoea agglomerans Pantoea vagans Phenotype Phosphates Photosynthesis Plant Development Plant Embryos Plant Leaves Plant Proteins Plant Roots Plants Polaromonas ginsengisoli Pseudoduganella violaceinigra Pseudomonas Pseudomonas fluorescens Rahnella Rahnella aquatilis Retention (Psychology) Rhodococcus erythropolis Rosa Salt Stress Sodium Chloride Sodium Chloride, Dietary Stenotrophomonas chelatiphaga Stenotrophomonas maltophilia Stenotrophomonas rhizophila Stenotrophomonas terrae Sterility, Reproductive Strains Technique, Dilution Threonine Triticum aestivum Tryptophan Tyrosine Vegetables Zea mays

Example 49

[Figure (not displayed)]

(7,8-Dichloro-4-(1H-pyrazol-4-yl)quinolin-2-yl)glycine was prepared essentially by the same method described above to prepare I-287.

The following compounds are prepared essentially by the same method described above to prepare I-177.

MS
I-#Starting MaterialStructure[M + 1]+
I-402[Figure (not displayed)]
[Figure (not displayed)]
[Figure (not displayed)]
407
I-180[Figure (not displayed)]
[Figure (not displayed)]
[Figure (not displayed)]
351
I-181[Figure (not displayed)]
[Figure (not displayed)]
[Figure (not displayed)]
351
I-265[Figure (not displayed)]
[Figure (not displayed)]
[Figure (not displayed)]
386
I-403[Figure (not displayed)]
[Figure (not displayed)]
[Figure (not displayed)]
319

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Patent 2024
Anabolism Glycine pyrazole

Example 80

[Figure (not displayed)]

Step 1: tert-Butyl 4-(2-(3-((2-(tert-butoxy)-2-oxoethyl)amino)propyl)-7,8-dichloroquinolin-4-yl)-1H-pyrazole-1-carboxylate. To a solution of tert-butyl 4-(7,8-dichloro-2-(3-oxopropyl)quinolin-4-yl)-1H-pyrazole-1-carboxylate (100 mg) and tert-butyl glycinate (62 mg) in DCM (0.5 mL) and EtOH (2.5 mL) were added NaBH3CN(100 mg) and acetic acid (1 drop). After stirring overnight, quenching with water, extraction with DCM, and concentration under reduced pressure, the residue was purified by a preparative thin layer chromatography (eluting with 15% MeOH in DCM) to afford the title compound (58 mg). MS: [M+1]+ 479.

Step 2: (3-(7,8-Dichloro-4-(1H-pyrazol-4-yl)quinolin-2-yl)propyl)glycine. To a solution of tert-butyl 4-(2-(3-((2-(tert-butoxy)-2-oxoethyl)amino)propyl)-7,8-dichloroquinolin-4-yl)-1H-pyrazole-1-carboxylate (15 mg) in DCM (3 mL) was added TFA (0.5 mL). The resultant solution was stirred over 6 hours at room temperature. After evaporation under reduced pressure, the residue was purified by preparative HPLC to afford the title compound (6.4 mg) (MS: [M+1]+ 379.1). 1H NMR (400 MHz, CD3OD): δ 8.179-8.202 (d, J=9.2 Hz, 1H), 8.140 (s, 1H), 7.690-7.713 (d, J=9.2 Hz, 1H), 7.596 (s, 1H), 4.019 (S, 2H), 3.324-3.365 (m, 4H), 2.338-2.391 (m, 2H) ppm

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Patent 2024
1H NMR Acetic Acid Anabolism Ethanol Glycine High-Performance Liquid Chromatographies Pressure pyrazole tert-butoxy tert-butyl glycinate TERT protein, human Thin Layer Chromatography

Example 7

The interaction of the chimeric anti-FOLR1 mAbs and FOLR1 was measured by Surface Plasmon Resonance (SPR) (Biacore 8k, GE Healthcare). Briefly, an anti-FOLR1 mAb was immobilized to a protein A sensor chip (GE Healthcare, Cat #: 29-1275-56) in HBS-P buffer (0.01 M HEPES pH 7.4, 0.15 M NaCl, 0.05% v/v Surfactant P20) with an injection flow rate at 10 μL/ml. The recombinant FOLR1 (Novoprotein, Cat #: C784) at variant concentrations was loaded with the flow rate of 30 μL/min in HBS-P buffer. Following antigen loading, the surface was regenerated with 10 mM glycine-HCl (pH 1.5). Sensor grams were fit with a 1:1 binding model using Biacore 8k evaluation software (GE Healthcare). The results of the binding affinity for the anti-FOLR1 mAbs are shown in Table 7.

TABLE 7
KD values for the anti-FOLR1 mAbs from a Biacore assay
Chimeric mAbKD (nM)
F40.547
F50.352
F72.33
F80.721
F100.704
F170.265
F1911.8
F201.00

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Patent 2024
Antigens Biological Assay Buffers Chimera DNA Chips FOLR1 protein, human Glycine HEPES Sodium Chloride Staphylococcal Protein A Surface-Active Agents Surface Plasmon Resonance

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Glycine is a colorless, crystalline amino acid that is used as a raw material in the production of various pharmaceutical and chemical products. It serves as a key component in buffer solutions and is commonly employed in the preparation of cell culture media and various biological assays.
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Tris-Glycine gels are a type of polyacrylamide gel used for electrophoresis, a technique for separating and analyzing proteins. These gels consist of a Tris-Glycine buffer system and are commonly used in SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) applications.
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PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.
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Protease inhibitor cocktail is a laboratory reagent used to inhibit the activity of proteases, which are enzymes that break down proteins. It is commonly used in protein extraction and purification procedures to prevent protein degradation.
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Nitrocellulose membranes are a type of laboratory equipment designed for use in protein detection and analysis techniques. These membranes serve as a support matrix for the immobilization of proteins, enabling various downstream applications such as Western blotting, dot blotting, and immunodetection.
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Triton X-100 is a non-ionic surfactant commonly used in various laboratory applications. It functions as a detergent and solubilizing agent, facilitating the solubilization and extraction of proteins and other biomolecules from biological samples.
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