Gossypium , chemistry

Latest Paper:

Genetika. 2007 Apr ;43 (4):508-15 17555127

[Species-specific features of the protein patterns of diploid cotton

seeds with A and D genomes and of some amphidiploids]

Sh Iu Iunuskhanov , D Iu Ataev , I Zh Kurbanbaev

Differences between species were revealed in electrophoretic patterns of

seed proteins of various diploid cotton species with A and D genomes and

some amphidiploids. Reference spectra and electrophoretic formulas were

compiled for representatives of diploid and amphidiploid species, and the

electrophoretic spectra were visually evaluated. They would allow

identification of various cotton species, varieties, and lines. Homology

between cotton species was estimated from the results of electrophoretic

protein studies. The homology between species of a single genome group

was shown to be closer than between species belonging to different

genome groups.

Mesh-terms: Diploidy; Genome, Plant; Gossypium, chemistry;

Gossypium, genetics; Plant Proteins, analysis; Plant Proteins, genetics;

Seeds, chemistry; Seeds, genetics; Species Specificity;

Most cited papers:

Anal Biochem. 1994 Nov 15;223 (1):7-12 7535022 [Cited: 24]

A modified hot borate method significantly enhances the yield of

high-quality RNA from cotton (Gossypium hirsutum L.).

C Y Wan , T A Wilkins

The isolation of biologically active RNA from cotton (Gossypium

hirsutum L.) is difficult due to interference by high levels of endogenous

phenolics, polysaccharides, and secondary metabolites. A modified hot

borate procedure was developed to combat these cellular constituents

during tissue homogenization, resulting in the quantitative recovery of

RNA suitable for hybridization analysis, in vitro translation, and cDNA

synthesis. The efficacy of several hot borate buffer adjuvants for the

qualitative and quantitative recovery of leaf RNA was monitored by

absorbance spectra, gel electrophoresis, protein, and cDNA synthesis. Of

the buffer adjuvants evaluated, polyvinylpyrrolidone-40 (PVP-40)

exhibited the single, most significant impact on the yield and quality of

RNA isolated from cotton leaves, although inclusion of deoxycholate

and/or Nonident-40 (NP-40) further enhanced the quality of the RNA.

The unsurpassed qualitative and quantitative recovery of total RNA from

cotton by hot borate buffer at alkaline pH, supplemented with PVP-40,

deoxycholate, and/or NP-40 had also proven satisfactory for other

recalcitrant plant species as well as for especially difficult tissue types.

Mesh-terms: Boric Acids; DNA, Complementary, biosynthesis;

Gossypium, chemistry; RNA, isolation & purification; Support, Non-U.S.

Gov't; Translation, Genetic;

Biochim Biophys Acta. 2001 Jan 12;1544 (1-2):196-206 11341929

[Cited: 5]

Isolation and characterization of a D-7 LEA protein from pollen that

stabilizes glasses in vitro.

W F Wolkers , S McCready , W F Brandt , G G Lindsey , F A Hoekstra

A heat-soluble protein present in substantial quantities in Typha latifolia

pollen was purified to homogeneity. The protein was subjected to

cyanogen bromide cleavage, and the peptides produced were separated by

HPLC chromatography and sequenced. The two sequences determined

were found to be related to the putative D76 LEA protein from Brassica

napus seeds and one of them to the D-7 LEA protein from upland cotton.

This suggests the pollen protein to be a member of the LEA group III

family of proteins. The secondary structure of the protein in solution and

in the dry state was investigated using Fourier transform IR spectroscopy.

Whereas the protein in solution was highly unordered, being largely in a

random coil conformation, the conformation was largely alpha-helical

after fast drying. Slow drying reversibly led to both alpha-helical and

intermolecular extended beta-sheet structures. When dried in the presence

of sucrose, the protein adopted alpha-helical conformation, irrespective of

drying rate. The effect of the protein on the stability of sucrose glasses

was also investigated. The dehydrated mixture of sucrose and the LEA

protein had higher glass transition temperatures and average strength of

hydrogen bonding than dehydrated sucrose alone. We suggest that LEA

proteins may play a role together with sugars in the formation of a tight

hydrogen bonding network in the dehydrating cytoplasm, thus conferring

long-term stability.

Mesh-terms: Amino Acid Sequence; Brassica, chemistry; Carbohydrate

Conformation; Electrophoresis, Polyacrylamide Gel; Glass; Gossypium,

chemistry; Plant Proteins, chemistry; Plant Proteins, isolation &

purification; Pollen, chemistry; Protein Structure, Secondary;

Spectroscopy, Fourier Transform Infrared; Sucrose, chemistry; Support,

Non-U.S. Gov't;

Plant Physiol. 1995 Aug ;108 (4):1691-701 7659756 [Cited: 5]

Solubilization and partial characterization of extensin fragments

from cell walls of cotton suspension cultures. Evidence for a covalent

cross-link between extensin and pectin.

X Qi , B X Behrens , P R West , A J Mort

Extensin, a major hydroxyproline (Hyp)-rich glycoprotein in walls of

cultured cells of dicotyledonous plants, is very difficult to solubilize. To

learn about the nature of the insolubilization, we have tested the ability of

a variety of selective hydrolytic methods, and combinations of them, to

liberate extensin or fragments of extensin from suspension-culture cell

walls. After the complete deglycosylation of cotton (Gossypium hirsutum

L.) walls, trypsinization solubilized 80% of the Hyp. The sequences of

three abundant peptides were: (a) serine-Hyp-Hyp-Hyp-Hyp-Hyp-Hyp-

serine-Hyp-Hyp-lysine, (b) serine-Hyp-Hyp-Hyp-Hyp-valine-lysine, and

(c) serine-Hyp-Hyp-serine-alanine-Hyp-lysine. After a sequential

treatment of walls with endopolygalacturonase, cellulase, -73 degrees C

anhydrous hydrogen fluoride solvolysis, and ammonium bicarbonate

extraction, only sugars indicative of rhamnogalacturonan I and protein

remained insoluble. Trypsin treatment of this residue liberated 50% of the

Hyp. A significant proportion of rhamnogalacturonan-associated sugars

co-solubilized and co-purified along with the extensin fragments

following the trypsinization. By sodium dodecyl sulfate gel

electrophoresis and gel filtration, the glycopeptides fell into two classes.

One class contained distinctly sized molecules with relative molecular

weights in the range of 4,000 to 24,000. The other class did not enter the

resolving gel and was hetero-disperse. After complete deglycosylation by

a 0 degrees C anhydrous hydrogen fluoride treatment, the first class was

little affected in its electrophoretic mobility, whereas the larger

heterogeneous material mostly entered the separating gel. After further

trypsinization of the deglycosylated peptides and analysis by capillary

zone electrophoresis, the peptides in both size classes were shown to

contain the sequences described above. From our observations we suggest

that cotton extensin becomes insolubilized into cell walls in part by

pectin-protein cross-links in addition to the protein-protein (or protein-

phenolic-protein) cross-links that have been repeatedly suggested.

Mesh-terms: Amino Acid Sequence; Amino Acids, analysis;

Carbohydrates, analysis; Cell Wall, chemistry; Cells, Cultured;

Glycoproteins, chemistry; Gossypium, chemistry; Hydroxyproline,

analysis; Molecular Sequence Data; Pectins, chemistry; Peptide

Fragments, chemistry; Plant Proteins, chemistry; Sequence Analysis;

Solubility; Spectrometry, Mass, Secondary Ion; Support, Non-U.S. Gov't;

Support, U.S. Gov't, Non-P.H.S.;

Biochim Biophys Acta. 1997 Apr 10;1351 (3):305-12 9130594

[Cited: 3]

Identification of a cotton fiber-specific acyl carrier protein cDNA by

differential display.

P Song , R D Allen

Transcripts from immature fibers and stripped ovules (fibers removed) of

cotton (Gossypium hirsutum L.) were compared by differential display to

identify cDNA fragments that represent mRNAs that are expressed

primarily in cotton fibers. Eight independent fiber-specific cDNA

fragments were isolated. One of these cDNAs had strong sequence

similarity with acyl carrier protein (ACP). A full-length cDNA for the

cotton fiber-specific ACP was isolated using a PCR cDNA library

screening technique. This 713 bp cDNA has an open reading frame that

encodes a 136 amino acid polypeptide. Overall nucleotide and amino acid

sequence identities with other plant ACP gene sequences averaged 66%

and 60% respectively. A 19 amino acid sequence surrounding the

prosthetic group attachment site is nearly identical to other plant ACP

genes. Northern blot analyses showed that transcripts homologous to this

fiber-specific ACP cDNA were predominantly expressed during the

elongation stage of fiber development. Initial genomic Southern blot

analysis indicated that a single copy of the fiber-specific ACP gene may

be present in both the cotton A and D genomes, since diploid Gossypium

species with A or D genomes gave identical bands. We speculate that this

putative fiber-specific ACP may play an important role in rapidly

elongating cotton fibers by contributing to the synthesis of membrane

lipids. It is also apparent that during the evolution of cotton a member of

the ACP gene family has been recruited for specific expression in cotton

fibers.

Mesh-terms: Acyl Carrier Protein, genetics; Acyl Carrier Protein,

metabolism; Amino Acid Sequence; Blotting, Northern; Blotting,

Southern; DNA, Complementary, genetics; Gene Expression Regulation,

Plant; Gene Library; Genetic Techniques; Gossypium, chemistry;

Gossypium, genetics; Gossypium, growth & development; Molecular

Sequence Data; Plant Proteins, genetics; Plant Proteins, metabolism;

Polymerase Chain Reaction, methods; RNA, Messenger, biosynthesis;

RNA, Plant, chemistry; RNA, Plant, genetics; Sequence Analysis, DNA;

Tissue Distribution;

Plant Physiol. 1995 Sep ;109 (1):269-75 7480326 [Cited: 3]

N-acylphosphatidylethanolamine in dry and imbibing cottonseeds.

Amounts, molecular species, and enzymatic synthesis.

J A Sandoval , Z H Huang , D C Garrett , D A Gage , K D Chapman

N-Acylphosphatidylethanolamine (NAPE), an unusual acylated derivative

of phosphatidylethanolamine (PE), was recently shown to be synthesized

from PE and free fatty acids in cotyledons of cotton (Gossypium hirsutum

L.) seedlings (K.D. Chapman, T.S. Moore [1993] Plant Physiol 102: 761-

769). Here we report that NAPE is present in dry seeds of cotton and

increases with time of imbibition from 2.31 nmol/seed in dry seeds to

4.26 nmol/seed in 4-h-soaked seeds. Total phospholipid/seed also

increased such that the relative percentage of NAPE was similar in dry

and soaked seeds (2.3 mol% compared to 2.6 mol%, respectively). The

major molecular species of NAPE were identified in both dry and soaked

seeds by fast atom bombardment mass spectrometry and collisionally

activated dissociation tandem mass spectrometry as 16:0/18:2-PE(N-

palmitoyl), 16:0/18:2-PE(N-linoleoyl), and 18:2/18:2-PE(N-palmitoyl).

The specific activity of NAPE synthase in seed extracts increased with

increasing time of imbibition from 35 pmol h-1 mg-1 protein in dry seeds

to 129 pmol h-1 mg-1 protein in 4-h-soaked seeds. Collectively, our

results indicate that NAPE is present in dry cottonseeds and synthesized

during imbibition. The biosynthesis of NAPE provides a mechanism for

maintaining membrane integrity during seed rehydration and may indicate

that NAPE plays a protective role in intracellular membranes of plant

tissues, as has been suggested for intracellular membranes of animal

tissues.

Mesh-terms: Acyltransferases, metabolism; Gossypium, chemistry;

Gossypium, metabolism; Kinetics; Molecular Structure;

Phosphatidylethanolamines, analysis; Phosphatidylethanolamines,

biosynthesis; Phosphatidylethanolamines, chemistry; Seeds, chemistry;

Seeds, metabolism; Spectrometry, Mass, Fast Atom Bombardment;

Support, Non-U.S. Gov't; Support, U.S. Gov't, Non-P.H.S.; Support, U.S.

Gov't, P.H.S.; Water;

J Exp Bot. 2002 Feb ;53 (367):323-31 11807136 [Cited: 2]

Plant allocation to defensive compounds: interactions between

elevated CO(2) and nitrogen in transgenic cotton plants.

Carlos E Coviella , Robert D Stipanovic , John T Trumble

Plant allocation to defensive compounds in response to growth in elevated

atmospheric CO(2) in combination with two levels of nitrogen was

examined. The aim was to discover if allocation patterns of transgenic

plants containing genes for defensive chemicals which had not evolved in

the species would respond as predicted by the Carbon Nutrient Balance

(CNB) hypothesis. Cotton plants (Gossypium hirsutum L.) were sown

inside 12 environmental chambers. Six of them were maintained at an

elevated CO(2) level of 900 micromol mol(-1) and the other six at the

current level of approximately 370 micromol mol(-1). Half the plants in

each chamber were from a transgenic line producing Bacillus

thuringiensis (Bt) toxin and the others were from a near isogenic line

without the Bt gene. The allocation to total phenolics, condensed tannins,

and gossypol and related terpenoid aldehydes was measured. All the

treatments were bioassayed against a non-target insect herbivore found on

cotton, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae). Plants had

lower N concentrations and higher C:N ratios when grown in elevated

CO(2). Carbon defensive compounds increased in elevated CO(2), low N

availability or both. The increase in these compounds in elevated CO(2)

and low N, adversely affected growth and survival of S. exigua. The

production of the nitrogen-based toxin was affected by an interaction

between CO(2) and N; elevated CO(2) decreased N allocation to Bt, but

the reduction was largely alleviated by the addition of nitrogen. The CNB

hypothesis accurately predicted only some of the results, and may require

revision. These data indicate that for the future expected elevated CO(2)

concentrations, plant allocation to defensive compounds will be affected

enough to impact plant-herbivore interactions.

Mesh-terms: Adaptation, Physiological; Animals; Bacillus thuringiensis,

pathogenicity; Bacterial Toxins, metabolism; Biological Assay; Carbon

Dioxide, metabolism; Carbon, metabolism; Gossypium, chemistry;

Gossypium, metabolism; Gossypium, parasitology; Gossypol,

biosynthesis; Host-Parasite Relations; Insecticides, metabolism; Nitrogen,

metabolism; Pest Control, Biological; Phenols, metabolism; Plant

Diseases, parasitology; Plant Leaves, chemistry; Plant Leaves,

metabolism; Plant Leaves, parasitology; Plants, Genetically Modified;

Protein Precursors, metabolism; Spodoptera, growth & development;

Support, Non-U.S. Gov't; Tannins, biosynthesis; Time Factors;

Plant Mol Biol. 1997 May ;34 (1):111-8 9177317 [Cited: 2]

Identification of a delta-TIP cDNA clone and determination of

related A and D genome subfamilies in Gossypium species.

D L Ferguson , R B Turley , R H Kloth

Tonoplast intrinsic proteins (TIPs) have been implicated in the process of

cell elongation, such as occurs in the developing cotton fiber. We have

isolated a cDNA clone (997 bp in length) from a cotton (Gossypium

hirsutum L.) library which putatively encodes a protein of 248 residues

(Mr 25079) with 85% identity to Arabidopsis delta-TIP. The derived

amino acid sequence included two conserved sequences associated with

major intrinsic proteins (SGxHxNPA at residues 78 to 85, NPA residues

at 197 to 199) and a cysteine residue at 116 which is reported to bind

mercury in Arabidopsis delta-TIP. The polymerase chain reaction was

used to generate partial genomic clones of the cotton delta-TIP. In

comparison to other genomic TIP sequences, the number (two) and

position of the introns were conserved in cotton. Comparing the TIP

sequences from cotton revealed two subfamilies, which were consistently

distinguished by a Tsp45I restriction site polymorphism. This

polymorphism was used to demonstrate that TIP subfamilies were

specific to either the A or D genomes of Gossypium. When delta-TIP

DNA fragments were amplified from cDNA of fiber 14 days after

anthesis, the A and D were found, indicating the presence of delta-TIP

transcripts in these elongating cells.

Mesh-terms: Amino Acid Sequence; Aquaporins; Arabidopsis Proteins;

Base Sequence; Cloning, Molecular; DNA, Complementary, isolation &

purification; DNA, Complementary, metabolism; DNA, Plant, chemistry;

DNA, Plant, genetics; Genome, Plant; Gossypium, chemistry;

Gossypium, genetics; Molecular Sequence Data; Multigene Family; Plant

Proteins, classification; Plant Proteins, genetics; Polymerase Chain

Reaction; Porins, classification; Porins, genetics;

Protoplasma. 2003 Jun ;221 (3-4):175-84 12802624 [Cited: 1]

Localization of sucrose synthase and callose in freeze-substituted

secondary-wall-stage cotton fibers.

Vadim V Salnikov , Mark J Grimson , Robert W Seagull , Candace H

Haigler

Methods for cryogenic fixation, freeze substitution, and embedding were

developed to preserve the cellular structure and protein localization of

secondary-wall-stage cotton (Gossypium hirsutum L.) fibers accurately

for the first time. Perturbation by specimen handling was minimized by

freezing fibers still attached to a seed fragment within 2 min after removal

of seeds from a boll still attached to the plant. These methods revealed

native ultrastructure, including numerous active Golgi bodies,

multivesicular bodies, and proplastids. Immunolocalization in the context

of accurate structure was accomplished after freeze substitution in acetone

only. Quantitation of immunolabeling identified sucrose synthase both

near the cortical microtubules and plasma membrane and in a proximal

exoplasmic zone about 0.2 microm thick. Immunolabeling also showed

that callose (beta-1,3-glucan) was codistributed with sucrose synthase

within this exoplasmic zone. Similar results were obtained from cultured

cotton fibers. The distribution of sucrose synthase is consistent with its

having a dual role in cellulose and callose synthesis in secondary-wall-

stage cotton fibers.

Mesh-terms: Cell Wall, chemistry; Cell Wall, ultrastructure; Cellulose,

biosynthesis; Cotton Fiber; Cryopreservation; Glucans, analysis;

Glucosyltransferases, analysis; Gossypium, chemistry; Gossypium,

enzymology; Immunohistochemistry; Microscopy, Electron; Polymers,

analysis; Support, Non-U.S. Gov't; Support, U.S. Gov't, Non-P.H.S.;

J Agric Food Chem. 2002 Nov 20;50 (24):7017-21 12428953 [Cited:

1]

Toxicity of (+)- and (-)-gossypol to the plant pathogen, Rhizoctonia

solani.

Lorraine S Puckhaber , Michael K Dowd , Robert D Stipanovic , Charles

R Howell

The dimeric sesquiterpene gossypol occurs naturally in cottonseed and

other parts of the cotton plant. Gossypol exists as enantiomers because of

the restricted rotation around the central binaphthyl bond. The (-)-

enantiomer is toxic to nonruminant animals while the (+)-enantiomer

exhibits little, if any, toxicity to these animals. Developing cotton plants

with low levels of the (-)-gossypol could expand the use of cottonseed as

a feed source. Gossypol also may play a role in protecting the plant from

pathogens. The relative toxicity of (+)- and (-)-gossypol to plant

pathogens has not been reported. We measured the concentration of (+)-

and (-)-gossypol in roots from cotton seedlings that were treated with the

biocontrol agent Trichoderma virens that induces biosynthesis of gossypol

and related terpenoids in cotton roots. (-)-Gossypol was the minor

enantiomer in control and treated roots, but levels were slightly higher in

roots from T. virens-treated seed. We also determined the toxicity of the

gossypol enantiomers and the racemate to the seedling disease pathogen

Rhizoctonia solani. The (+)- and (-)-enantiomers of gossypol and the

racemate are equally effective in inhibiting growth of this pathogen. The

lethal doses of the gossypols required to kill the pathogen appeared to be

similar, but their toxicities are significantly less than those of related

cotton and kenaf sesquiterpenes. The results indicate that altering the

enantiomeric ratio in cotton roots will not adversely affect the resistance

of seedlings to the seedling pathogen R. solani.

Mesh-terms: Gossypium, chemistry; Gossypium, microbiology;

Gossypol, analysis; Gossypol, pharmacology; Plant Diseases,

microbiology; Plant Roots, chemistry; Rhizoctonia, drug effects;

Rhizoctonia, growth & development; Seedling, chemistry; Seedling,

microbiology; Seeds, chemistry; Stereoisomerism; Trichoderma;

Plant Physiol. 2001 Nov ;127 (3):1234-42 11706202 [Cited: 1]

Sucrose phosphate synthase activity rises in correlation with high-

rate cellulose synthesis in three heterotrophic systems.

V M Babb , C H Haigler

Based on work with cotton fibers, a particulate form of sucrose (Suc)

synthase was proposed to support secondary wall cellulose synthesis by

degrading Suc to fructose and UDP-glucose. The model proposed that

UDP-glucose was then channeled to cellulose synthase in the plasma

membrane, and it implies that Suc availability in cellulose sink cells

would affect the rate of cellulose synthesis. Therefore, if cellulose sink

cells could synthesize Suc and/or had the capacity to recycle the fructose

released by Suc synthase back to Suc, cellulose synthesis might be

supported. The capacity of cellulose sink cells to synthesize Suc was

tested by analyzing the Suc phosphate synthase (SPS) activity of three

heterotrophic systems with cellulose-rich secondary walls. SPS is a

primary regulator of the Suc synthesis rate in leaves and some Suc-

storing, heterotrophic organs, but its activity has not been previously

correlated with cellulose synthesis. Two systems analyzed, cultured

mesophyll cells of Zinnia elegans L. var. Envy and etiolated hypocotyls

of kidney beans (Phaseolus vulgaris), contained differentiating tracheary

elements. Cotton (Gossypium hirsutum L. cv Acala SJ-1) fibers were also

analyzed during primary and secondary wall synthesis. SPS activity rose

in all three systems during periods of maximum cellulose deposition

within secondary walls. The Z. elegans culture system was manipulated to

establish a tight linkage between the timing of tracheary element

differentiation and rising SPS activity and to show that SPS activity did

not depend on the availability of starch for degradation. The significance

of these findings in regard to directing metabolic flux toward cellulose

will be discussed.

Mesh-terms: Asteraceae, chemistry; Asteraceae, metabolism; Cell

Differentiation; Cell Wall, metabolism; Cells, Cultured; Cellulose,

biosynthesis; Cellulose, chemistry; Fructose, metabolism;

Glucosyltransferases, metabolism; Gossypium, chemistry; Gossypium,

metabolism; Hypocotyl, metabolism; Models, Molecular; Phaseolus,

chemistry; Phaseolus, metabolism; Plant Leaves, cytology; Plant Leaves,

metabolism; Starch, metabolism; Sucrose, metabolism; Support, Non-

U.S. Gov't; Uridine Diphosphate Glucose, metabolism;

Chem Senses. 2008 Jan 8; : 18184638

Sensory Deafferentation Transsynaptically Alters Neuronal GluR1

Expression in the External Plexiform Layer of the Adult Mouse Main

Olfactory Bulb.

Kathryn A Hamilton , Stephanie Parrish-Aungst , Frank L Margolis ,

Ferenc Erdélyi , Gabor Szabó , Adam C Puche

Altered distribution of the alpha-amino-3-hydroxy-5-methylisoxazole-4-

propionic acid (AMPA) receptor subunit GluR1 has been linked to

stimulation-dependent changes in synaptic efficacy, including long-term

potentiation and depression. The main olfactory bulb (OB) remains plastic

throughout life; how GluR1 may be involved in this plasticity is

unknown. We have previously shown that neonatal naris occlusion

reduces numbers of interneuron cell bodies that are immunoreactive for

GluR1 in the external plexiform layer (EPL) of the adult mouse OB. Here,

we show that immunoreactivity of mouse EPL interneurons for GluR1 is

also dramatically reduced following olfactory deafferentation in

adulthood. We further show that expression of glutamic acid

decarboxylase (GAD) 65, 1 of 2 GAD isoforms expressed by adult

gamma-aminobutyric acidergic interneurons, is reduced, but to a much

smaller extent, and that in double-labeled cells, immunoreactivity for the

Ca(2+)-binding protein parvalbumin (PV) is also reduced. In addition,

GluR1 expression is reduced in presumptive tufted cells and interneurons

that are negative for GAD65 and PV. Consistent with previous reports,

sensory deafferentation resulted in little neuronal degeneration in the

adult EPL, indicating that these differences were not likely due to death of

EPL neurons. Together, these results suggest that olfactory input regulates

expression of the GluR1 AMPA receptor subunit by tufted cells that may

in turn regulate GluR1 expression by interneurons within the OB EPL.

Ann Intern Med. 1949 Aug ;31 (2):216-27 18136037

Pulmonary disease manifestations of ankylosing spondylarthritis.

K A HAMILTON

Bioorg Med Chem Lett. 2007 Sep 7; : 17900896

Synthesis and evaluation of substituted benzoisoquinolinones as

potent inhibitors of Chk1 kinase.

Robert M Garbaccio , Shaei Huang , Edward S Tasber , Mark E Fraley ,

Youwei Yan , Sanjeev Munshi , Mari Ikuta , Lawrence Kuo , Constanine

Kreatsoulas , Steve Stirdivant , Bob Drakas , Keith Rickert , Eileen S

Walsh , Kelly A Hamilton , Carolyn A Buser , James Hardwick , Xianzhi

Mao , Stephen C Beck , Marc T Abrams , Weikang Tao , Rob Lobell ,

Laura Sepp-Lorenzino , George D Hartman

From HTS lead 1, a novel benzoisoquinolinone class of ATP-competitive

Chk1 inhibitors was devised and synthesized via a photochemical route.

Using X-ray crystallography as a guide, potency was rapidly enhanced

through the installation of a tethered basic amine designed to interact with

an acidic residue (Glu91) in the enzyme pocket. Further SAR was

explored at the solvent front and near to the H1 pocket and resulted in the

discovery of low MW, sub-nanomolar inhibitors of Chk1.

J Neurophysiol. 2007 Jan 10; : 17215500

Group I Metabotropic Glutamate Receptors Are Differentially

Expressed by Two Populations of Olfactory Bulb Granule Cells.

Thomas Heinbockel , Kathryn A Hamilton , Matthew Ennis

In the main olfactory bulb, several populations of granule cells (GCs) can

be distinguished based on the soma location either superficially,

interspersed with mitral cells within the mitral cell layer (MCL), or

deeper, within the GC layer (GCL). Little is known about the

physiological properties of superficial GCs (sGCs) vs. deep GCs (dGCs).

Here, we used patch-clamp recording methods to explore the role of

Group I metabotropic glutamate receptors (mGluRs) in regulating the

activity of GCs in slices from wildtype and mGluR -/- mutant mice. In

wildtype mice, bath application of the selective Group I mGluR agonist

DHPG depolarized and increased the firing rate of both GC subtypes. In

the presence of blockers of fast synaptic transmission (APV, CNQX,

gabazine), DHPG directly depolarized both GC subtypes. The two GC

subtypes responded differentially to DHPG in mGluR1-/- and mGluR5-/-

mice, however. DHPG depolarized sGCs in slices from mGluR5-/- mice,

but it had no effect on sGCs in slices from mGluR1-/- mice. By contrast,

DHPG depolarized dGCs in slices from mGluR1-/- mice, but it had no

effect on dGCs in slices from mGluR5-/- mice. Previous studies have

shown that mitral cells express mGluR1, but not mGluR5. The present

results therefore suggest that sGCs are more similar to mitral cells than

dGCs in terms of mGluR expression.

Trends Microbiol. 2006 Feb 3; : 16460942

Animal movements and the spread of infectious diseases.

Eric M Fèvre , Barend M de C Bronsvoort , Katie A Hamilton , Sarah

Cleaveland

Domestic and wild animal population movements are important in the

spread of disease. There are many recent examples of disease spread that

have occurred as a result of intentional movements of livestock or

wildlife. Understanding the volume of these movements and the risks

associated with them is fundamental in elucidating the epidemiology of

these diseases, some of which might entail zoonotic risks. The importance

of the worldwide animal trade is reviewed and the role of the unregulated

trade in animals is highlighted. A range of key examples are discussed in

which animal movements have resulted in the introduction of pathogens

to previously disease-free areas. Measures based on heightened

surveillance are proposed that mitigate the risks of new pathogen

introductions.

Neuroscience. 2005 ;133:819-29 15896912

Properties of external plexiform layer interneurons in mouse

olfactory bulb slices.

K A Hamilton , T Heinbockel , M Ennis , G Szabó , F Erdélyi , A Hayar

In the external plexiform layer (EPL) of the main olfactory bulb, apical

dendrites of inhibitory granule cells form large numbers of synapses with

mitral and tufted (M/T) cells, which regulate the spread of activity along

the M/T cell dendrites. The EPL also contains intrinsic interneurons, the

functions of which are unknown. In the present study, recordings were

obtained from cell bodies in the EPL of mouse olfactory bulb slices.

Biocytin-filling confirmed that the recorded cells included interneurons,

tufted cells, and astrocytes. The interneurons had fine, varicose dendrites,

and those located superficially bridged the EPL space below several

adjacent glomeruli. Interneuron activity was characterized by high

frequency spontaneous excitatory postsynaptic potential/currents that

were blocked by the alpha-amino-3-hydroxy-5-methylisoxazole-4-

propionic acid (AMPA)/kainate receptor antagonist 6-cyano-7-

nitroquinoxaline-2,3-dione and largely eliminated by the voltage-sensitive

Na+ channel blocker, tetrodotoxin. Interneuron activity differed markedly

from that of tufted cells, which usually exhibited spontaneous action

potential bursts. The interneurons produced few action potentials

spontaneously, but often produced them in response to depolarization

and/or olfactory nerve (ON) stimulation. The responses to depolarization

resembled responses of late- and fast-spiking interneurons found in other

cortical regions. The latency and variability of the ON-evoked responses

were indicative of polysynaptic input. Interneurons expressing green

fluorescent protein under control of the mouse glutamic acid

decarboxylase 65 promoter exhibited identical properties, providing

evidence that the EPL interneurons are GABAergic. Together, these

results suggest that EPL interneurons are excited by M/T cells via

AMPA/kainate receptors and may in turn inhibit M/T cells within spatial

domains that are topographically related to several adjacent glomeruli.

Mesh-terms: Action Potentials, physiology; Animals; Cell Shape,

physiology; Excitatory Postsynaptic Potentials, physiology; Green

Fluorescent Proteins, genetics; Interneurons, cytology; Interneurons,

physiology; Mice; Mice, Inbred C57BL; Mice, Transgenic; Olfactory

Bulb, cytology; Olfactory Bulb, physiology; Organ Culture Techniques;

Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't;

Research Support, U.S. Gov't, P.H.S.;

J Agric Food Chem. 2004 Nov 17;52 (23):6969-76 15537305

Bollgard II cotton: compositional analysis and feeding studies of

cottonseed from insect-protected cotton (Gossypium hirsutum L.)

producing the Cry1Ac and Cry2Ab2 proteins.

Kathryn A Hamilton , Paul D Pyla , Matthew Breeze , Tammy Olson ,

Menghe Li , Edwin Robinson , Sean P Gallagher , Roy Sorbet , Yin Chen

Bollgard II cotton event 15985 producing the Cry1Ac and Cry2Ab2

proteins has been developed by genetic modification to broaden the

spectrum of insects to which the plant is tolerant and to provide an insect

resistance management tool to impede the onset of resistance. The

purpose of this study was to evaluate the composition and nutrition of

Bollgard II cotton, relative to the use for food and animal feed, compared

to that of conventional cotton varieties. Compositional analyses were

conducted to measure proximate, fiber, amino acid, fatty acid, gossypol,

and mineral contents of cottonseed from a total of 14 U.S. field sites over

two years. Compositional analysis results showed that the cottonseed and

cottonseed oil from Bollgard II cotton were comparable in their

composition to those of the conventional control cotton line and other

commercial varieties. The composition data are supported by nutritional

safety studies conducted with dairy cows, catfish, and quail. Results from

these studies showed that Bollgard II performed similarly to the

conventional control cotton varieties. These data demonstrate that

Bollgard II cotton is compositionally and nutritionally equivalent to

conventional cotton varieties. These data support the conclusion that

Bollgard II cotton is as safe and nutritious as conventional cotton for food

and feed use.

Mesh-terms: Amino Acids, analysis; Animal Feed; Animals; Bacterial

Proteins, genetics; Bacterial Toxins, genetics; Cattle; Cottonseed Oil,

chemistry; Dietary Fiber, analysis; Endotoxins, genetics; Fatty Acids,

analysis; Gossypium, chemistry; Gossypium, genetics; Gossypol,

analysis; Ictaluridae; Minerals, analysis; Plants, Genetically Modified,

chemistry; Plants, Genetically Modified, genetics; Quail;

Can Nurse. 1959 May ;55 (5):409-13 13652065

Emotional problems of the worker.

K A HAMILTON

Mesh-terms: Emotions; Nursing; Occupational Health;

Gossypium spp.

Cotton

Useful references 400, 452The cotton-seed consists of two parts: the hull, from which the staple cotton lint and linters arise, and the kernel, from which the oil and meal are obtained. The nutritive value of cottonseed products depends on proportions of husks and lint. 

The husk is sometimes separated from the kernel before

crushing, but often the whole seed is extracted for oil.

Undecorticated oilcake is much richer in fibre and lower in

protein. The term "Egyptian cotton cake" refers to the

undecorticated cake of black seeds, and "Bombay cotton

cake" is the term used for the undecorticated cake of white

seeds. The cotton fibres of white seeds cover the whole

surface and are very difficult to remove; if the cottonseed

cake is broken, the fibres can be seen. 

For every ton of lint in seed cotton there are approximately

1.7 tons of cottonseed. One ton of seed yields about 200 kg

of oil, 500 kg of cottonseed meal and 300 kg of hulls. The

residual oil in hydraulic-press cake is usually between 4%

and 8%, in screw-press cake between 3% and 5%, and in

solvent-extracted meal less than 3%. 

 

GOSSYPOL. The seed embryo contains innumerable

glands filled with a yellow pigment called gossypol.

McDonald et al. (553) state that cotton seeds may contain

from 0.3-20 g/kg DM of gossypol, and concentrations of 4-

17 g/kg DM have been quoted for the kernels. Gossypol is

a polyphenolic aldehyde which is an antioxidant and

polymerisation inhibitor and is toxic to monogastric

animals: pigs and rabbits are the most sensitive, whereas

poultry are more tolerant. The general symptoms of

gossypol toxicity are constipation, depressed appetite and

loss of weight; death usually results from circulatory

failure. Although acute toxicity is low, ingestion of a small

amounts over a prolonged period can be lethal. It is

important to distinguish between free (soluble in 70-30 v/v

aqueous acetone) and bound gossypol since only the

former is considered to be physiologically active. 

The free gossypol content of cottonseed meal decreases

during processing and varies according to the methods

used. In new seed, free gossypol accounts for 0.4-1.4% of

the weight of the kernel. Screw pressed materials have

200-500 mg free gossypol/kg, pre- 

pressed solvent extracted meals 200-700 and solvent

extracted 

1000-5000 mg/kg. Processing conditions have to be

carefully controlled to prevent loss of protein quality

owing to binding of gossypol to lysine at high

temperatures. Fortunately the shearing effect of the screw

press in the expeller process is an efficient gossypol

inactivator at temperatures which do not reduce protein

quality. 

It is generally considered that pig and poultry diets should

not contain more than 100 mg free gossypol/kg and that

inclusions of cottonseed meal should be between 50-100

kg/t of feed. Particular care is required with laying hens

since comparatively low levels of the meal may cause an

olive green discoloration of the yolk in storage. An

associated pink discoloration of the albumen is now

considered to be due to cyclopropenoids and not gossypol

as was once thought. Treatment with ferrous sulphate can

ameliorate the biological effects of gossypol; the amount

of iron to be added is largely empirical. To reduce the

effects of gossypol on growth, the following proportions of

iron to free gossypol have been used: for cattle 1:1, for

broilers 2:1, for layers 4:l and for pigs 1:1. 

Mature ruminant animals do not show ill-effects even

when they consume large quantities of cottonseed meal but

young cattle are much more susceptible to its toxic effects. 

USES 

COTTONSEED MEAL or COTTONSEED CAKE. 

Cottonseed meal is an excellent protein supplement for

cattle. The limitations on effective utilization of this

product in rations for swine and poultry are of minor

significance for ruminant animals. 

Both decorticated and undecorticated cottonseed meal

have a constipating effect on cattle, which is beneficial in

feeds with a high molasses content. Calves are susceptible

to the harmful effects of gossypol because of incomplete

rumen development; hence it is recommended that

concentrates for calves under five months of age contain

no more than 10-15% cottonseed meal. 

Cottonseed meal has a relatively low rumen degradability

and is therefore a good source of by-pass protein and

especially useful in rations for milking cows. 

With Creole and crossbred village cows in Mauritius

(normal milk yield 4-5 litres), Boodoo et al. (573) reported

that producers obtained over 9 litres milk/day over 300

days with peak milk yield at 12 litres/day, by

supplementing sugarcane tops and roadside grasses with

0.25 kg cottonseed cake/litre milk. The results were the

same as those obtained with 0.5 kg/litre of commercial

compound feed (17% CP). 

Cottonseed meal can be used safely and profitably in pig

rations. Rations containing up to 0.01% free gossypol can

be used without the addition of iron salts. On the basis of a

typical analysis this means that the safe upper limit for a

good screw-press or prepress solvent meal is about 20%

and for direct solvent-extracted meal about 5%. Over these

levels, iron must be added in a 1:1 weight ratio to free

gossypol. 

Cottonseed meal can also be used in rations for growing

chickens if the free gossypol does not exceed 0.03%.

Below this level, iron (2:1 iron to free gossypol) will

completely overcome the depressing effect. As cottonseed

is low in lysine, it may be necessary to make up for this

deficiency by adding lysine. Prepress solvent-extracted

meal may be used in layer feed if the gossypol has been

inactivated with iron (4:1 iron to free gossypol), if the free

gossypol content is not above 0.4%. 

As determination of the amount of free gossypol in a meal

requires laboratory facilities, ferrous sulphate may

routinely be added to diets containing cottonseed meal. For

instance, for layers 0.05% iron (corresponding to 0.25%

ferrous sulphate septahydrate) can routinely be included in

diets containing up to 10% cottonseed meal and up to

0.16% iron in rations containing greater amounts. Broilers

cannot tolerate more than 0.07% iron in the diet. 

WHOLE COTTONSEED 

Whole cottonseed may be used as a feed for mature cattle,

as is often done where proper milling equipment is not

available. It is usually soaked in water and fed in small

quantities as a supplement to green feed. 

Compared to the extracted meal, it has a lower protein

content and a much higher oil content. High levels may

cause scouring. The degradability characteristics of the

protein should be similar. Delinting of FUZZY WHOLE

COTTONSEED to DELINTED WHOLE COTTONSEED

results in a higher protein and fat content and lower fibre

content. 

Neill et al. (574) report the use of up to 20-30% WCS in

intensive finishing rations for Brahman-cross steers.

Although OM digestibility declined, intake was unaffected

and LWG maintained 0.86 kg/day. They advise that WCS

can be used without detriment to at least 20% in finishing

diets, as a total feed during drought and for male animals

(rams) without affecting libido. No evidence of gossypol

toxicity was found but they suggest caution when feeding

WCS to young stock. 

By treating with 5% ferrous sulphate solution, it can safely

be included in small quantities in pig rations. 

COTTONSEED HULLS 

Cottonseed hulls are the parts of the cottonseed head that is

left after the cotton and meal have been extracted and are

normally considered to be a very low quality feed for

ruminants. The results of two recent trials have shown that

they are capable of supporting moderate growth rates in

sheep. When a small amount of by-pass protein was added

to the diet of cottonseed hulls + urea + 50 g lucerne +

vitamins/minerals, the growth rate of lambs exceeded 130

g/day (567) and wool growth was increased from 6 to 9

g/day. 

Investigation of the rumens of these animals showed that

protozoa were either eliminated or in very low population

densities. This could be the reason why cottonseed hulls

support such reasonable growth rates, even without

supplementation with by-pass protein. Intake of cottonseed

hulls by sheep is higher (c. 1 kg DM/day) than would be

expected of a 40% digestible feed and this is possibly

associated with a rapid breakdown of the indigestible

material in the rumen (567). 

Norbaev (568) reports problems of hepatosis associated

with the feeding of cottonseed husks in Russia. Meat

inspection of 1004 Karakul sheep slaughtered in

Uzbekistan revealed hepatosis in 41%, cirrhosis in 10.7%

and multiple abscesses of the liver in 3.7%. In a feeding

experiment on 30 sheep, in which the daily ration

comprised 2 kg cotton husks, 0.5 kg oatmeal and 0.2 kg

lucerne hay. Liver disease developed within 2 months.

Values for blood albumin and glucose were low. 

As cotton hulls are available at the mills where the meal is

produced, mixtures of meal and hulls can be bought. An

economical fattening ration for cattle is 20% meal and

80% hulls, together with 3-4 kg of grass daily and a

mineral supplement. 

Various experiments with dairy cows cited by Coppock et

al. (569) show that the roughage value of CSH is better

than grass hays and nearly equal to high quality

bermudagrass but inferior to mixed clover hay. There is no

effect when CSH is substituted for corn or barley. There is

some evidence that CSH fibre maintains milk and fat yield

even when DM digestibility of the diet declines as a result

of substitution of CSH for lucerne hay (570). In some

diets, CSH may have a special value because of a

stimulatory effect on feed intake, not through improvement

in digestibility. 

Cottonseed hulls have frequently been included in

complete diets for young, growing and lactating cattle.

Replacement of rice bran with up to 20% CSH did not

effect the milk yield or fat content of Murrah buffaloes

(571). 

One novel application is reported by Yang et al. (572) of

the feeding of cottonseed hulls to pigs in China, after use

as a growth medium for 4 harvests of mushrooms

(Pleurotus sp.). The optimum dose was 10% of the diet

mixed with concentrates, which gave the same growth rate

and carcass quality as a control diet. 

COTTON WOOD 

Cotton wood is also used in some countries as a cattle

fodder. Stems, branches and leaves can be ground, ensiled

or hydrolysed with sodium hydroxide for feeding to beef

cattle. 

 

As % of dry matter

DM CP CF Ash EE NFECa P Ref

Whole seeds, India

94.320.621.54.9 20.132.9 378

Whole seeds, Egypt

91.221.523.25.5 26.223.6 512

Bombay cotton cake

87.723.024.76.6 5.5 40.2 "

Egyptian cotton cake 87.926.424.26.6 5.7 37.1 "

Oilcake with hulls, mechanically extr-acted, Israel 91.426.924.05.0 6.8 37.3 0.22 0.63 365

Oilcake with hulls, mechanically extr-acted, Uganda 88.026.225.66.2 5.7 36.3 69

Oilcake with hulls, mechanically extr-acted, Zimbabwe 94.530.57.9 7.0 7.6 47.0 499

Oilcake without hulls, mechanically extracted, Israel 92.347.712.56.6 5.4 27.8 0.22 1.34 365

Oil meal without hulls, solventextracted, Iraq 94.340.315.76.8 0.6 36.6 0.31 1.11 182

Oil meal without hulls, prepresssolvent extracted, USA 89.946.115.17.1 0.7 31.0 0.17 1.36 452

Cottonseed hulls, West Indies 86.213.930.03.6 8.8 43.7 191

Cotton wood, Venezuela 5.3 64.98.1 0.9 20.8 537

Digestibility (%)

Animal CP CF EE NFEME Ref

Seeds, Indian variety Cattle 68.421.691.2 22.2 2.46 378

Seeds, Egyptian variety

Sheep 68.475.986.6 49.8 3.45 512

Bombay cotton cake

Sheep 77.220.393.8 53.7 2.13 "

Cake with hulls, expressed Sheep 75.038.095.0 66.0 2.59 365

Cake with hulls, expressed Cattle 72.057.091.3 80.4 3.02 499

Cake without hulls, expressed Sheep 83.045.097.0 74.0 3.06 365

Hulls Sheep 0.0 49.078.0 43.0 1.74 "

Nylon bag degradability

a b c 12hr 48hrRef(%) (%) (/hour) (%) (%)

Whole cotton seed, Ghana

DM 21.253.00.0320 62.8 627

Cotton seed, Korea

DM 27.141.70.0398 60.6 "

Cotton hulls, Brazil DM 2.8 51.00.0098 13.0 "[P (rumen degradability at time t) = a+b*(1-exp(-c*t))]

Amino acid composition as % of crude

protein

Oilcake Ref 452

Arg Cys GlyHisIls Leu Lys Met Phe Thr Try Tyr Val11.1 1.5 4.1 2.6 3.2 5.9 4.1 1.3 5.4 3.2 1.1 2.7 4.5

References69, 182, 191, 365, 378, 400, 452, 499, 512, 537, 553, 567, 568, 569, 570, 571, 572, 573, 574, 627

AbstractsBuffaloes(232), Cattle(77), Cattle(108), Cattle(179), Cattle(181), Cattle(200), Cattle(275), Cattle(283), Cattle(330), Cattle(333), Cattle(334), Cattle(380), Cattle(444), Cattle(476), Cattle(589), Cattle(630), Cattle(656), Cattle(683), Composition(156), Composition(186), Composition(207), Composition(476), Dairy(83), Dairy(218), Dairy(220), Dairy(319), Dairy(618), Degradability(419), Goats(38), Goats(212), Guinea-pigs(347), NaOH treatment(266), Pigs(73), Pigs(75), Pigs(144), Pigs(183), Pigs(462), Poultry(111), Poultry(149), Poultry(196), Poultry(217), Poultry(261), Poultry(318), Poultry(349), Poultry(423), Poultry(424), Poultry(531), Rabbits(57), Rabbits(216), Rabbits(221), Rabbits(347), Sheep(96), Sheep(212), Sheep(271), Sheep(441), Sheep(457), Sheep(682), Silage(108), Steam treatment(266), Tannin treatment(457), Toxicity(169), Toxicity(217), Toxicity(237), Toxicity(465), Turkeys(422)

Other papers by authors:

2594117April, 1952

Bonotto 260/123.5

Prepared cotton seed material for solvent extraction and processes for preparation and solvent extraction of cotton seed materials

3579496May, 1971

Martinez et al.

260/123.5

3615657October, 1971

Gastrock et al.

260/123.5PROCESS FOR PRODUCING COTTONSEED PROTEIN CONCENTRATE

3814748June, 1974

Olson et al.

260/123.5