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PubMed2004

Effects of soybean Kunitz trypsin inhibitor on the cotton boll weevil (Anthonomus grandis).

The cotton boll weevil, Anthonomus grandis, is an economically important pest of cotton in tropical and subtropical areas of several countries in the Americas, causing severe losses due to their damage in cotton floral buds. Enzymatic assays using gut extracts from larval and adult boll weevil have demonstrated the presence of digestive serine proteinase-like activities. Furthermore, in vitro assays showed that soybean Kunitz trypsin inhibitor (SKTI) was able to inhibit these enzymes. Previously, in vivo effects of black-eyed pea trypsin chymotrypsin inhibitor (BTCI) have been demonstrated towards the boll weevil pest. Here, when neonate larvae were reared on an artificial diet containing SKTI at three different concentrations, a reduction of larval weight of up to 64% was observed for highest SKTI concentration 500 microM. The presence of SKTI caused an increase in mortality and severe deformities of larvae, pupae and adult insects. This work therefore represents the first observation of a Kunitz trypsin inhibitor active in vivo and in vitro against A. grandis. Bioassays suggested that SKTI could be used as a tool in engineering crop plants, which might exhibit increased resistance against cotton boll weevil.

Animal Feed↗
PubMed2004

Terpenoid aldehyde formation and lysigenous gland storage sites in cotton: variant with mature glands but suppressed levels of terpenoid aldehydes.

A new cotton variant with reduced levels of terpenoid aldehydes (sesquiterpenoids and sesterterpenoids (heliocides)) was isolated from the progeny of hemizygous cotton (Gossypium hirsutum cv. Coker 312) transformed with antisense (+)-delta-cadinene synthase cDNA. Southern analysis of leaf DNA digested with HindIII, Pst or KpnI restriction endonucleases did not detect any antisense cdn1-C1 DNA in the genome of the variant. The gossypol content in the seed of the variant was markedly lower than in the seed of T1 antisense plants. Eighty-nine percent of the variant seed had a 71.1% reduction in gossypol and the foliage of the variant plants showed a 70% reduction in gossypol and a 31% reduction in heliocides. Compared to non-transformed plants there was no reduction in the number of lysigenous glands in the seed of the variant. The cotton variant shows uncoupling of terpenoid aldehyde synthesis and gland formation. The cotton variant may have resulted from somaclonal variation occurring in the callus tissue during the transformation-regeneration process.

Aldehydes↗
PubMed2002

A beta-tubulin-like cDNA expressed specifically in elongating cotton fibers induces longitudinal growth of fission yeast.

Using cDNA Representational Difference Analysis (RDA) techniques, we isolated a cDNA that was expressed specifically in cotton fibers but not in the ovules of a fuzzless-lintless mutant (fl). We designated it as Gh-BTubL for it shares high sequence identity with known plant and yeast beta-tubulins. RT-PCR and robotic cDNA dot blot analyses indicated that the expression of Gh-BTubL was correlated with the elongation pattern of cotton fibers. In situ hybridization results verified that there was no Gh-BTubL mRNA in fl ovules while it was easily detected in the elongating wild type cotton fiber cells. Overexpression of Gh-BTubL in fission yeast induced longitudinal growth of the host cells by 1.74-fold, with no apparent effect on other aspects of the host cells. We suggest that Gh-BTubL plays an important role in cotton fiber elongation and we believe that elucidation of the control mechanisms for expression of tubulin-like proteins may help improve fiber quality and productivity.

Amino Acid Sequence↗
PubMed1999

Identification of the bright-greenish-yellow-fluorescence (BGY-F) compound on cotton lint associated with aflatoxin contamination in cottonseed.

In order to characterize the structure of the bright-greenish-yellow-fluorescence (BGY-F) compound on cotton lint associated with aflatoxin contamination in cotton seed, various in vitro and in vivo natural BGY-F reaction products were prepared. Under similar high pressure liquid chromatography separation with variable wavelength and programmable fluorescence detection (HPLC-UV/FL), combined with atmospheric pressure ionization and mass spectral determinations it was found that the BGY-F reaction products prepared from three preparations: (a) kojic acid (KA) + peroxidase (soybean peroxide or horseradish type VI and type II) + H2O2, or (b) detached fresh cotton locules + KA + H2O2, or (c) attached field cotton locules that were treated with a spore suspension of aflatoxigenic Aspergillus flavus, all resulted in identical chromatographic characteristics, and all exhibited a molecular weight of 282. Further characterization of the BGY-F reaction product with 1H- and 13C-NMR spectroscopic analysis revealed that it was a dehydrogenator dimer of 2 KA, linked through the C-6 positions.

Aflatoxins↗
PubMed1995

Cotton-wool spots and the early diagnosis of giant cell arteritis.

BACKGROUND: Giant cell arteritis is a common cause of severe visual loss in older individuals. Patients often present to the ophthalmologist having already lost vision in one eye. Detection of early ophthalmoscopic signs that precede irreversible visual loss in giant cell arteritis would allow preventative treatment in an otherwise frequently blinding disease. METHODS: Case presentations. RESULTS: Seven patients with mild visual symptoms and results of an ophthalmologic examination significant for cotton-wool spots were found to have giant cell arteritis. On specific questioning, six of seven patients described constitutional symptoms consistent with giant cell arteritis. Six patients had an abnormally elevated Westergren erythrocyte sedimentation rate. Temporal artery biopsy confirmed giant cell arteritis in six patients. The seventh patient received a diagnosis of polymyalgia rheumatica. Prompt treatment with corticosteroids led to preservation of vision and uneventful resolution of the cotton-wool spots in all seven patients. CONCLUSION: Cotton-wool spots are an early ophthalmoscopic finding in giant cell arteritis and can precede severe visual loss. Recognition of the significance of cotton-wool spots, use of laboratory studies, and prompt treatment may preserve vision in an otherwise frequently blinding disease.

Aged↗
PubMed1995

Retinal distortion and cotton-wool spots associated with epiretinal membrane contraction.

PURPOSE AND BACKGROUND: Several articles on surgery for macular pucker have noted the presence of intraretinal white spots, presumably cotton-wool spots, associated with epiretinal membranes. It was proposed that membrane contraction and resultant nerve fiber layer distortion might lead to blockage of axoplasmic flow and thus a cotton-wool spot. To see whether the observations supported this concept, the authors studied patients with epiretinal membranes associated with intraretinal white spots involving the center of the fovea, where there are normally no retinal vessels and the only axons are in Henle fiber layer. METHODS: Four patients with epiretinal membranes associated with prominent areas of intraretinal whitening involving the center of the fovea were examined before and after surgery. Observations and measurements were made from projected fundus photographs. RESULTS: The intraretinal white spots, which appeared preoperatively to lie in the center of the fovea, proved to have been pulled in from more peripheral retina by the epiretinal membrane. These white areas were associated with surprisingly great distortion of the inner retina. Stretching of the inner retina seemed to play a larger role in the retinal distortion than full-thickness retinal folding. CONCLUSION: The appearance and time course of these intraretinal white lesions were consistent with cotton-wool spots caused by mechanical disruption of axonal flow. The marked retinal stretching and distortion of the innermost layers of the retina were the most striking findings in this study and also seemed consistent with the concept that membrane contraction and resultant nerve fiber layer distortion might lead to blockage of axoplasmic flow and thus a cotton-wool spot.

Axonal Transport↗
PubMed2001

Enzymes and chelating agent in cotton pretreatment.

Desized cotton fabric and cotton seed-coat fragments (impurities) have been treated with commercial cellulase (Celluclast 1.5 L), hemicellulase-pectinase (Viscozyme 120 L) and xylanase (Pulpzyme HC) enzymes. Seed-coat fragments hydrolyzed much faster than the cotton fabric itself. This relative difference in hydrolysis rates makes possible a direct enzymatic removal of seed-coat fragments from desized cotton fabric. Addition of chelating agents such as ethylenediamine-tetra-acetic acid (EDTA) markedly enhanced the directed enzyme action. Pretreatments carried out in acidic solution at pH 5 increased the lightness of seed-coat fragments, contrary to the samples treated in neutral medium at pH 7. Alkaline scouring resulted in darker seed-coat fragments except for the samples pretreated with Pulpzyme HC plus EDTA. This effect is similar to that observed in the biobleaching process in pulp and paper industry.

Calcium↗
PubMed2000

Genetic variability and molecular responses of root penetration in cotton.

Compacted soils restrict root penetration hindering productivity. In this paper, genetic variability of cotton (Gossipium spp.) root capacity to penetrate hard soil layers and the patterns of gene expression during penetration event were investigated. To mimic hard soil layers, wax-petrolatum mixtures were used. Genetic variability among 27 cotton genotypes for the root capacity to penetrate wax-petrolatum disks of 500-700 g wax/kg of mixture was high indicating that breeding efforts targeted to improve this trait can be successful. In the root tips of a cotton strain with high root penetrating ability (G. hirsutum HS 200) which penetrated through wax-petrolatum disks (P), quantity of four polypeptides with molecular weights 35-66 kDa increased compared to those root tips which grew in the absence of mechanical impedance (NP). Differential display showed significant differences in the sets of mRNA expressed in P and NP roots. Out of a total of 917 cDNAs scored in the differential display experiment, 118 cDNAs, or 13%, were specific to P roots and hence could be associated with the root penetration event. Further detailed study of gene expression in penetrated roots will pinpoint molecular factors involved in root penetration ability in cotton.

Journal Article↗
PubMed2000

Use of cotton rats for preclinical evaluation of measles vaccines.

The continued prevalence and medical impact of measles worldwide has created interest in the development of new generations of measles vaccines. Monkeys can be used for preclinical testing of these vaccines. However, a more practical and less expensive animal model is highly desirable, particularly for initial vaccine development and evaluation. Cotton rats have been shown to support the replication of different strains of measles virus (MV), and thus may be useful for these purposes. To test this concept, the immunogenicity and protective efficacy of two standard (Moraten and trivalent measles, mumps, rubella) and four experimental (two recombinant ALVAC, one ISCOM subunit and live attenuated Edmonston-Zagreb) MV vaccines were evaluated in naïve cotton rats, and cotton rats with passively acquired MV-specific neutralizing serum antibodies. All of the test vaccines were immunogenic and protected naíve animals from pulmonary infection and viral dissemination. However, under the conditions utilized, only the Edmonston-Zagreb vaccine provided such protection to animals with significant levels of passively acquired MV-specific neutralizing antibodies. The results of these tests and the potential of using cotton rats as an animal model for preliminary testing of MV vaccines are discussed.

Animals↗
PubMed2003

Hidden health costs of pesticide use in Zimbabwe's smallholder cotton growers.

Balancing the numerous benefits that may accrue from pesticide use on cotton, farmers face health hazards. Pesticide-induced acute symptoms significantly increased the cost-of-illness in a survey of 280 smallholder cotton growers in two districts of Zimbabwe. Cotton growers lost a mean of 180 Zimbabwe dollars in Sanyati and 316 Zimbabwe dollars per year in Chipinge on pesticide-related direct and indirect acute health effects. These values are equivalent to 45% and 83% of annual household pesticide expenditures in the two districts. The time spent recuperating from illnesses attributed to pesticides averaged 2 days in Sanyati and 4 days in Chipinge during the 1998/1999 growing season. These pesticide health cost estimates represent lower bounds only; they omit chronic pesticide health effects as well as suffering and other non-monetary costs. Acute pesticide symptoms were determined in large part by pesticide use practices, notably the lack of protective clothing. Yet many smallholder farmers misunderstood pesticide health hazards, and so did little to protect themselves. Despite the use of simple color codes, 22% of smallholder cotton growers in Sanyati and 58% in Chipinge did not know how the four colored triangles communicated increasing degrees of pesticide toxicity. Better farmer education in exposure averting strategies is needed. Likewise, fuller accounting for hidden health costs in future would allow farmers to make more informed decisions about agricultural pest management.

Agricultural Workers' Diseases↗
PubMed2000

The mechanism of cellulase action on cotton fibers: evidence from atomic force microscopy.

Two cellulases from Trichoderma reesei--an exoglucanase, CBH I, and an endoglucanase, EG II--alone and in combination were incubated with cotton fibers. The effects of the cellulases on the surfaces of the cotton fibers were examined by atomic force microscopy. At high magnification, the physical effects on the fibers caused by the two types of enzymes were considerably different. Treatment with CBH I resulted in the appearance of distinct pathways or tracks along the length of the macrofibril. Treatment with EG II appeared to cause peeling and smoothing of the fiber surface. In combination, their effect was observed to be greatest when both enzymes were present simultaneously. When fibers smoothed by treatment with EG II were treated subsequently with CBH I, further evidence of path way formation caused by the action of CBH I along the fibers was observed. Incubation with a cellulase from Thermotoga maritima that lacks a cellulose binding domain had no effect on the surface of cotton fibers. These images provide the first physical evidence of differences in the effect of cellulase components action on the surface of cotton fibers and provide evidence for the movement or tracking of CBH I along the fibers. The first AFM image of CBH I molecules are presented.

Cellulase↗
PubMed2000

Cloning, expression and purification of recombinant cotton rat interleukin-5.

The coding sequence of the hispid cotton rat (Sigmodon hispidus) interleukin-5 (IL-5) was isolated by a combination of reverse transcription (RT)-PCR and RACE protocols from concanavalin A stimulated spleen cells. The open reading frame of 399 bp encodes a polypeptide of 132 amino acids. Comparison with the rat, mouse, gerbil and human counterparts revealed 88, 88, 87 and 75% identity at the nucleotide level and 88, 90, 89 and 70% at the amino acid level, respectively. The entire coding sequence, minus the putative signal peptide sequence, was inserted into an inducible Escherichia coli expression vector. The recombinant protein possessed an expected molecular mass of 14kDa and was located in bacterial inclusion bodies. A purification scheme under reducing and denaturing conditions followed by subsequent successive dialysis steps led to the recovery of a recombinant dimeric cotton rat IL-5. The biological activity of the recombinant protein was demonstrated in a murine cell line proliferation assay. This activity was specifically inhibited by rat monoclonal antibodies directed against mouse IL-5. Together with specific antibodies that can be generated easily, cotton rat IL-5 constitutes a useful tool for extending the use of the cotton rat animal model in the study of various human pathogens.

Amino Acid Sequence↗
PubMed1999

Cloning, expression and purification of recombinant cotton rat interferon-gamma.

The hispid cotton rat (Sigmodon hispidus) has proven to be an excellent small animal model; however, immunological studies have been limited due to a lack of available reagents. We report cloning of the cotton rat interferon-gamma (IFN-gamma) cDNA from concanavalin A-stimulated spleen cells using a combination of reverse transcription polymerase amplification reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) protocols. The open reading frame of 513 nucleotides encodes a 170 amino acid (aa) protein followed by a stop codon with a predicted molecular mass of 19548Da. Cotton rat IFN-gamma shares 63, 60, 43 and 43% identity with the hamster, gerbil, mouse and rat counterpart, respectively. IFN-gamma nucleotide sequence corresponding to aa 18-153 was expressed in Escherichia coli under tryptophan promoter control, either fused to a single initiating codon or fused to the thioredoxin coding sequence. Both expression products were found exclusively in bacterial inclusion bodies. Two purification schemes have been developed to purify the product fused to a single methionine. One of them is fast and leads to the recovery of a pure product suitable for use in antibody production. The second protocol, which includes chromatographic steps, allows the use of the purified product for in vitro demonstration of biological activity in a viral cytopathic reduction assay on cotton rat cells.

Amino Acid Sequence↗
PubMed2006

Genetic diversity of source germplasm of Upland cotton in China as determined by SSR marker analysis.

The genetic diversity of 43 sources of Upland cotton germplasm with different parental origins, breeding periods, and ecological growing areas in China were studied on the basis of simple sequence repeat (SSR) markers. A total of 130 gene alleles with 80% polymorphism were detected from 36 SSR primers. The number of alleles per primer ranged from two to eight with an average of 3.6. The polymorphism information content (PIC) range was 0.278-0.865, with an average of 0.62. The average genotype diversity index (H') was 1.102, the highest was 2.039 and the lowest was 0.451. The average coefficient of the genetic similarity of SSR markers among source germplasm was 0.610, ranging from 0.409 to 0.865. These indicated that the genetic diversity at the genomic level of the selected source germplasm was rich, and was representative of the diversity of the germplasms, in general. The diversity at the genome level of the base germplasm from the second and third breeding periods was decreased compared to that of the first period, indicating that the cotton genetic background in China became narrow gradually. The diversity of SSR markers among the base germplasm from early maturity cotton growing areas in the north was higher than those from the Huanghe and Yangtze growing areas. The molecular marker genetic similarity index of the domestic varieties was higher than that in the introduced varieties, which indicates that the genetic diversity in domestic cultivars was lower than that in the introduced varieties. This study gives an overview of the genetic diversity of the cotton germplasm base in China, and provides a guide for breeders to develop new cultivars efficiently.

China↗
PubMed1982

The host-parasite relationships in normal and protein-malnourished cotton rats infected with Litomosoides carinii (Nematoda: Filarioidea).

A total of 120 young cotton rats were fed ad libitum for 10 weeks on either synthetic diets containing 2.5, 5, 10 or 15% casein or on a stock breeding diet containing 20% crude protein. Half of the animals in each group were infected with the filarial nematode Litomosoides carinii and the food intake and body weight of each animal was measured. There was a trend towards increased food consumption in protein-deficient cotton rats and this was increased further in infected animals; however, differences between groups were not significant. Low protein diets reduced the rate of growth of uninfected and infected cotton rats; filarial infection intensified these adverse effects of under-nutrition and increased the number of rats which died in the lowest protein group. Protein deficiency had little effect on the number of L. carinii which developed, although fewer parasites became established in cotton rats fed on a 2.5% diet than in those fed on a 10% diet. Parasites developing in protein-deficient animals were shorter than those developing in well-fed ones and embryogenesis was retarded in female worms from protein-deficient animals. The onset of patency was retarded in protein-deficient animals and the microfilaraemia which developed subsequently was lower up to 120 days post-infection.

Animals↗
PubMed1995

The host-parasite relationships in pyridoxine (vitamin B6) deficient cotton rats infected with Litomosoides carinii (Nematoda: Filaroidea).

This paper demonstrates that the establishment and growth of the filarial nematode parasite, Litomosoides carinii, is reduced in pyridoxine-deficient cotton rats. Young cotton rats were assigned to one of three dietary: vitamin B6-deficient cotton rats (B6-AL) were fed a pyridoxine-free diet ad libitum; pair-fed controls (B6 + PF) were fed the same amount of pyridoxine-free diet as animals in the deficient group and given daily oral supplements of 100 micrograms pyridoxine; and pyridoxine-sufficient controls (B6 + AL) were fed the pyridoxine-free diet ad libitum and supplemented daily with 100 micrograms pyridoxine. Half of each group was infected with 50 L3 of L. carinii by subcutaneous injection 8 weeks after the start of the experimental feeding period. B6-deficient cotton rats ate less (P < 0.001) and gained less weight (P < 0.001) than B6-supplemented controls. The levels of microfilaraemia in deficient animals, measured weekly throughout the experiment by taking blood smears, was significantly lower than in supplemented animals (P < 0.001). The deficient rats became latent for L. carinii at 20 weeks post-infection, whereas there was patent microfilaraemia in rats in the other dietary groups until the end of the experiment. Smaller (P < 0.001) and fewer (P < 0.05) adult worms were recovered from the pleural and abdominal cavities of deficient animals than from either pair-fed or sufficient controls at autopsy 28 week post-infection.

Animals↗
PubMedDate not supplied

Efficient production of L-(+)-lactic acid using mycelial cotton-like flocs of Rhizopus oryzae in an air-lift bioreactor.

L-(+)-Lactic acid production was enhanced in a culture of Rhizopus oryzae by induction of a mycelial flocs morphology. By conventional culture the morphology of R. oryzae is that of a pellet-like cake; however, when mineral support and poly(ethylene oxide) are added to the culture, the morphology of R. oryzae takes on a cotton-like appearance. The formation of these cotton-like mycelial flocs was induced by the addition of 5 ppm poly(ethylene oxide) into a 12-14 h culture containing 3 g/L of the mineral support before the formation of the conventional pellet morphology. The cotton-like flocs were also formed in cultures grown in an air-lift bioreactor. This morphology allowed effective mass transfer inside the flocs and effective fluidity of culture broth in an air-lift bioreactor. L-(+)-Lactic acid concentration produced by mycelial flocs in an air-lift bioreactor, with the support and poly(ethylene oxide), was 104.6 g/L with a yield of 0.87 using 120 g/L of glucose as the substrate; for this culture without both, the concentration was 43.2 g/L. These results demonstrate that cotton-like mycelial flocs are the optimal morphology for use in the air-lift bioreactor culture of R. oryzae.

Bioreactors↗
PubMed2000

Leaf senescence-like characteristics contribute to cotton's premature photosynthetic decline.

Leaf and canopy photosynthesis of cotton (Gossypium hirsutum L.) declines as the crop approaches cutout, just as the assimilate needs for reproductive growth are peaking. Our objective with this study was to determine whether this decline is due to remobilization of leaf components to support the reproductive growth or due to some cue from the changing environmental conditions during the growing season. Field studies were conducted in 1995-1996 at Stoneville, Mississippi, using six cotton genotypes and two planting dates (early and late), which produced two distinctly different cotton populations reaching cutout at different times. Among the six genotypes were a photoperiod sensitive line (non-flowering) and its counter part which had photoperiod insensitive genes backcrossed four times to the photoperiod sensitive line (flowering). This pair was used to assess the degree that the photosynthetic decline could be attributed to reproductive sink development. Leaf CO(2)-exchange rate (CER) and chlorophyll (Chl) fluorescence measurements were taken in mid-August, a period corresponding to cutout for the early planted plots, and those leaves were collected. Leaf Chl level, soluble protein level, various soluble carbohydrate levels and Rubisco activities were assayed on those leaves. Averaged across years, leaf CER and soluble protein levels were reduced approximately 14% and 18%, respectively, for the early planted compared to the late planted cotton. Neither leaf Chl levels or Chl fluorescence Fv/Fm values for Photosystem II yield were altered by the planting date. In 1996, leaves from the non-flowering line had 12% greater Chl and 20% greater soluble protein levels than the flowering line. However, in 1996, the CER of the early planted non-flowering line was reduced 10% compared to the late planted. Although remobilization of leaf N to reproductive growth appears to be the principle component causing the cutout photosynthetic decline, the data also indicate that environmental factors can play a small role in causing the decline.

Journal Article↗