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A longitudinal observation of early pulmonary responses to cotton dust.

AIMS: To examine early adverse pulmonary effects of exposure to cotton dust, and to identify potential risk factors, including atopy for pulmonary responses to cotton dust. METHODS: Spirometry, methacholine challenge testing, and questionnaire; performed among 101 non-smoking newly hired textile workers at baseline (prior to starting work), and at 3, 12, and 18 months after starting work. Concentrations of airborne cotton dust in various work areas were measured at each follow up survey using vertical elutriators. RESULTS: The incidence of non-specific respiratory symptoms was 8% at three months, then diminished afterwards. Substantial acute cross shift drops in FEV(1) at each follow up survey, and longitudinal declines in FVC and FEV(1) after 12 months of exposure were observed. Airway responsiveness to methacholine increased with follow up time, and was more pronounced among atopics. Increasing airway responsiveness was strongly correlated with cross shift drops in FEV(1). In addition, one or more respiratory symptoms at three months was significantly, and pre-existing atopy marginally significantly, associated with cross shift drops in FEV(1) after adjusting for other covariates and confounders. CONCLUSION: Results suggest that non-specific respiratory symptoms, decreasing lung function, and increasing airway responsiveness are early pulmonary responses to cotton dust. In addition, the occurrence of respiratory symptoms and increasing airway responsiveness, as well as atopy, may be important predictors for acute changes in lung function among cotton textile workers.

Adolescent↗

Composition and ethanol production potential of cotton gin residues.

Cotton gin residue (CGR) collected from five cotton gins was fractionated and characterized for summative composition. The major fractions of the CGR varied widely between cotton gins and consisted of clean lint (5-12%),hulls (16-48%), seeds (6-24%), motes (16-24%), and leaves (14-30%). The summative composition varied within and between cotton gins and consisted of ash (7.9-14.6%), acid-insoluble material (18-26%), xylan (4-15%),and cellulose (20-38%). Overlimed steam-exploded cotton gin waste was readily fermented to ethanol by Escherichia coli KO11. Ethanol yields were feedstock and severity dependent and ranged from 58 to 92.5% of the theoretical yields. The highest ethanol yield was 191 L (50 gal)/t, and the lowest was 120 L (32 gal)/t.

Agriculture↗

[Isolation by suppression-subtractive hybridization of genes preferentially expressed during early and late fiber development stages in cotton].

As a main natural fiber source, cotton plays an important role in human life. To identify genes preferentially expressed during early and late cotton fiber development, we constructed two fiber subtracted libraries on the basis of PCR-selected subtraction using a pool of nonfiber tissues as the same driver and 10 days postanthesis (DPA) and 20 DPA fiber cells as testers, respectively. Through differential screening, 292 clones in both libraries were identified as being preferentially expressed during fiber development. Sequence analysis showed that 31 unique sequences were found in the library of 10 DPA fibers and 48 unique sequences were obtained in the library of 20 DPA fibers. In addition, there were 13 unique clones in common in both libraries. Many previously reported cotton fiber-related genes were included in both libraries. Northern hybridization was performed to further confirm the differential expression and to determine the pattern of mRNA accumulation of selected clones. As a result, all selected cDNAs showed highly preferential expression in developing cotton fibers. Four genes-putative gibberellin-regulated protein, putative tonoplast intrinsic protein, putative plasma membrane intrinsic protein, and Gossypium hirsutum putative membrane protein-were identified in 10 DPA fiber subtracted library, and they were found to be expressed a lot during early fiber development. On the other hand, those genes screened out of 20 DPA fiber subtracted library, like arabinogalactan protein and fiber glycosyl hydrolase family 19 protein, were found highly expressed in fibers with the maximal transcription level during developmental switch from elongation to cellulose deposition. By subtraction between fibers and five nonfiber tissues, two sets of genes were identified, and their fiber-specific or fiber-preferential expression indicated that they are involved in the network that controls cotton fiber development.

Cloning, Molecular↗

Cotton dust contains proteolytic and elastolytic enzymes not inhibited by alpha-1-proteinase inhibitor.

Others have found proteolytic activity in cotton dust and correlated this with the prevalence of acute byssinosis. The present study characterized the proteases in aqueous extracts of cotton dust after controlling for potential artifact from microbial contamination. We tested cotton dust, generated in a model card room operated by the U.S. Department of Agriculture (USDA), for total proteolytic activity (hydrolysis of azocasein) and elastolytic activity (degradation of insoluble 3H-elastin). Both coarse dust and a respirable fraction (aerodynamic diameter less than or equal to 1.7 microgram) yielded proteolytic activity. Proteolysis was inhibited by EDTA (39%), diisopropylfluorophosphate (DFP) (33%) and alpha-1-proteinase inhibitor (alpha 1 PI) (30%), but not by iodoacetic acid or pepstatin. Elastolytic activity was also present, and was inhibited by DFP (80%) and alpha 1 PI (70%), but not by EDTA. We conclude that cotton dust contains preformed metallo- and serine proteases, and serine elastolytic protease(s) which are incompletely inhibited by alpha 1 PI. These findings may be relevant to the development of chronic lung disease in people exposed to cotton dust.

Byssinosis↗

Prevalence of byssinosis and respiratory symptoms among spinners in Sudanese cotton mills.

Byssinosis, other respiratory symptoms, and ventilatory capacity were investigated in 186 male spinners in two Sudanese textile mills located in Khartoum and Hassaheisa. The prevalence of byssinosis was 37% among Khartoum spinners where coarse cotton was processed, significantly higher (p less than 0.01) than the 1% found among Hassaheisa spinners, where fine cotton was used. The prevalence of chronic bronchitis was 29% in Khartoum and 2% in Hassaheisa spinners. A significant reduction in forced expiratory volumes (FEV1 after a shift) was observed in spinners of both factories. Our findings provide evidence that very low rates or absence of byssinosis, related respiratory symptoms, and significant functional abnormalities (FEV1) in the late stages of cotton yarn processing all depend on the quality of cotton used and low levels of cotton dust concentration.

Adolescent↗

Two-dimensional correlation spectroscopy and principal component analysis studies of temperature-dependent IR spectra of cotton-cellulose.

The FTIR spectra were measured for raw Uplands Sicala-V2 cotton fibers over a temperature range of 40-325 degrees C to explore the temperature-dependent changes in the hydrogen bonds of cellulose. These cotton-cellulose spectra exhibited complicated patterns in the 3800-2800 cm(-1) region and thus were analyzed by both the exploratory principal component analysis (PCA) and two-dimensional (2-D) correlation spectroscopy methods. The exploratory PCA showed that the spectra separate into two groups on the basis of thermal degradation of the cotton-cellulose and the consequent breakage of intersheet H-bonds present in its structure. Frequency variables, which strongly contributed to each principal component highlighted in its loadings plot, were linked to the frequencies assigned to vibrations of the OH groups involved in different kinds of H-bonds, as well as to vibrations of the CH groups. Deeper insights into reorganization of the temperature-dependent hydrogen bonding were obtained by 2-D correlation spectroscopy. Synchronous and asynchronous spectra were analyzed in the temperature ranges of 40 to 150 and 250 to 320 degrees C, the ranges indicated by PCA. Detailed band assignments of the OH stretching region and changes in the patterns of the hydrogen bonding network of the cotton-cellulose were proposed with the aid of the 2-D correlation spectroscopy analysis. Below 150 degrees C, distinctly different bands assigned to the less stable Ialpha and the more stable Ibeta interchain H-bonds O-6-H-6...O-3' were observed at about 3230 and 3270 cm(-1), respectively. Evaporation of water entrapped in the cellulose network was examined by means of the band at about 3610 cm(-1). The cooperativity of hydrogen bonds, which play a key role in the cellulose conformation, was monitored by frequencies assigned to intrachain H-bonds. It was possible to separate the frequencies assigned to the O-2-H-2...O-6 and O-3-H-3...O-5 intrachain H-bonds into two separate ranges, the spread of which was controlled by the cooperativity effect. The temperature dependence of the asynchronous spectra indicated that the less stable O-3-H-3...O-5 bonds gave rise to an absorption extending from 3300 to 3384 cm(-1), while the more stable O-2-H-2...O-6 bonds were characterized by the absorption between 3400 and 3470 cm(-1). The final breaking of the inter- and intrachain H-bonds, which occurs at the higher temperatures, was monitored by the asynchronous peaks at 3533 and 3590 cm(-1), respectively. On the basis of both the exploratory PCA and 2-D correlation spectroscopy investigations, it was possible to extract well-defined wavenumber ranges assigned to different kinds of intra- and interchain hydrogen bonds, as well as to the free OH groups of the cotton-cellulose.

Algorithms↗

An efficient protein preparation for proteomic analysis of developing cotton fibers by 2-DE.

Preparation of high-quality proteins from cotton fiber tissues is difficult due to high endogenous levels of polysaccharides, polyphenols, and other interfering compounds. To establish a routine procedure for the application of proteomic analysis to cotton fiber tissues, a new protocol for protein extraction was developed by optimizing a phenol extraction method combined with methanol/ammonium acetate precipitation. The protein extraction for 2-DE was remarkably improved by the combination of chemically and physically modified processes including polyvinylpolypyrrolidone (PVPP) addition, acetone cleaning, and SDS replacement. The protocol gave a higher protein yield and vastly greater resolution and spot intensity. The efficiency of this protocol and its feasibility in fiber proteomic study were demonstrated by comparison of the cotton fiber proteomes at two growth stages. Furthermore, ten protein spots changed significantly were identified by MS/tandem MS and their potential relationships to fiber development were discussed. To the best of our knowledge, this is the first time that a protocol for protein extraction from cotton fiber tissues appears to give satisfactory and reproductive 2-D protein profiles. The protocol is expected to accelerate the process of the proteomic study of cotton fibers and also to be applicable to other recalcitrant plant tissues.

Databases, Protein↗

Long-term selection for resistance to transgenic cotton expressing Bacillus thuringiensis toxin in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae).

Selection experiments for resistance to transgenic Bt cotton expressing Cry1Ac toxin in Helicoverpa armigera (Hübner) were conducted using a leaf-feeding method with 42 selection episodes over 45 generations. The cotton bollworm developed resistance to transgenic Bt cotton after 12 generations (F12) of selection. The survival rate of F12 neonates feeding on leaves of seedling stage for 4 days and boll-opening stage for 5 days of R19 line were ca 34 and 72%, respectively, compared with ca 0 and 40% for a non-selected sister strain (NYCS), but lower than or similar to that of F12 feeding on leaves of non-Bt cotton, Sumian 12 (ca 87 or 76%). Resistance to B thuringiensis HD-1 Dipel in neonates (F12) was ca 6-fold. After 42 generations of selection, the strain developed a very high level of resistance to Cry1Ac protoxin, 210 g kg(-1) MVPII wettable powder and 200 g litre(-1) MVPII liquid formulation, the resistance ratios being 1680-, 1780- and ca 1200-fold, respectively, compared with a laboratory susceptible strain (HZS). When compared with the non-selected NYCS, the resistance ratios to the above toxins were ca 540-, 580- and 510-fold, respectively, but to Dipel only ca 16-fold. The results indicated that it is very important to develop and implement effective resistance-management strategies and to detect early resistance to Bt cotton in field populations.

Animals↗

Airborne endotoxin concentrations in various work areas within a cotton mill in Central America.

Gram-negative bacteria and their endotoxins have been shown to cause profound changes in the structure and function of mammalian lungs. Airborne exposures in humans have resulted in bronchoconstriction and symptoms of chest tightness and dyspnea. Cotton dust is often heavily contaminated and endotoxin has been postulated to be the agent in cotton dust which is responsible for the byssinosis syndrome. Previous studies of cotton dust have revealed variable amounts of contamination by bacterial endotoxin, with the variability determined in large part by botanical and climatic characteristics. We report here the results of an environmental study conducted in a cotton mill located in the tropics of Central America using locally grown cotton. Results indicate that airborne endotoxin was present in all yarn preparation and weaving areas and ranged from a low of 18 EU/m3 in weaving to a high of 3138 EU/m3 in opening areas. Airborne levels and the amount of endotoxin per nanogram of dust were not higher than values obtained in temperate climates.

Air Pollutants, Occupational↗

Admicellar polymerization of styrene on cotton.

Thin-film coating on cotton by the admicellar polymerization process was investigated. In this work, styrene was used as the monomer to coat styrene on cotton. The effects of surfactant, styrene, initiator, and electrolyte concentrations on the polymerization process were determined. The polystyrene film formed was characterized by SEM, FTIR, and GPC. The increase in the hydrophobicity of the treated cotton surface was determined by the drop test. Results show that polystyrene thin film was successfully formed on cotton, resulting in cotton that can resist wetting by a water droplet for longer than 30 min.

Journal Article↗

Identification of dna components required for induction of cotton leaf curl disease.

Cotton leaf curl disease (CLCuD) is a major constraint to cotton production in Pakistan. Infectious clones of the monopartite begomovirus cotton leaf curl virus (CLCuV), associated with diseased cotton, are unable to induce typical symptoms in host plants. We have identified and isolated a single-stranded DNA molecule approximately 1350 nucleotides in length which, when coinoculated with the begomovirus to cotton, induces symptoms typical of CLCuD, including vein swelling, vein darkening, leaf curling, and enations. This molecule (termed DNA beta) requires the begomovirus for replication and encapsidation. The CLCuV/DNA 1/DNA beta complex, together with a similar complex previously identified in Ageratum conyzoides, represent members of an entirely new type of infectious, disease-causing agents. The implications of this finding to our understanding of the evolution of new disease-causing agents are discussed.

Base Sequence↗

Topographic changes in the optic disc in eyes with cotton-wool spots and primary open-angle glaucoma.

Changes in the topography of the optic disc in 26 eyes with cotton-wool spots displaying defects in the retinal nerve-fiber layer and in 31 eyes with early primary open-angle glaucoma showing a similar degree of such defects were studied by computer-assisted optic disc analyzer and then compared with 27 controls. Changes in the cup-to-disc ratio, cup volume, and ratio of rim area to disc area were not significant in eyes with cotton-wool spots. The quotient of cup volume divided by cup-to-disc ratio eyes with of primary open-angle glaucoma was greater than that in eyes with cotton-wool spots. For the detection of nerve loss in eyes with cotton-wool spots, the image analyzer, which identified the notches in the horizontally sectioned contour line of the cup, was more sensitive than stereoscopic detection of the notches in the rim (P less than 0.05). The image analyzer enabled the detection of slight nerve-fiber loss by examination of the contour line of the cup in eyes with cotton-wool spots.

Adult↗

Cleaved AFLP (cAFLP), a modified amplified fragment length polymorphism analysis for cotton.

In certain plant species including cotton (Gossypium hirsutum L. or Gossypium barbadense L.), the level of amplified fragment length polymorphism (AFLP) is relatively low, limiting its utilization in the development of genome-wide linkage maps. We propose the use of frequent restriction enzymes in combination with AFLP to cleave the AFLP fragments, called cleaved AFLP analysis (cAFLP). Using four Upland cotton genotypes (G. hirsutum) and three Pima cotton (G. barbadense), we demonstrated that cAFLP generated 67% and 132% more polymorphic markers than AFLP in Upland and Pima cotton, respectively. This resulted in 15.5 and 25.5 polymorphic cAFLP markers per AFLP primer combination, as compared to 9.1 and 11.0 polymorphic AFLP. The cAFLP-based genetic similarity (GS) is generally lower than the AFLP-based GS, even though both marker systems are overall congruent. In some cases, cAFLP can better resolve genetic relationships between genotypes, rendering a higher discriminatory power. Given the high-resolution power of capillary-based DNA sequencing system, we further propose that AFLP and cAFLP amplicons from the same primer combination can be pooled as one sample before electrophoresis. The combination produced an average of 18.5 and 31.0 polymorphic markers per primer pair in Upland and Pima cotton, respectively. Using several restriction enzyme combinations before pre-selective amplification in combination with various frequent 4 bp-cutters or 6 bp-cutters after selective amplification, the pooled AFLP and cAFLP will provide unlimited number of polymorphic markers for genome-wide mapping and fingerprinting.

Breeding↗

Resistance gene analogue markers are mapped to homeologous chromosomes in cultivated tetraploid cotton.

Degenerate primers designed from conserved motifs of known plant resistance gene products were used to amplify genomic DNA sequences from the root-knot nematode (Meloidogyne incognita) resistance genetic source, Upland cotton (Gossypium hirsutum) cultivar Auburn 634 RNR. A total of 165 clones were isolated, and sequence analysis revealed 57 of the clones to be novel nucleotide sequences, many containing the resistance (R)-protein nucleotide-binding site motif. A cluster analysis was performed with resistance gene analogue (RGA) nucleotide sequences isolated in this study, in addition to 99 cotton RGA nucleotide sequences already deposited in GenBank, to generate a phylogenetic tree of cotton R genes. The cotton RGA nucleotide sequences were arranged into 11 groups and 56 sub-groups, based on genetic distances. Multiple sequence alignments were performed on the RGA sequences of each sub-group, and either the consensus sequences or individual RGA sequences were used to design 61 RGA-sequence-tagged site primers. A recombinant inbred line (RIL) population of cultivated tetraploid cotton was genotyped using RGA-specific primers that amplified polymorphic fragments between the two RIL parents. Nine RGA markers were mapped to homeologous chromosomes 12 and 26, based on linkage to existing markers that are located on these chromosomes.

Amino Acid Sequence↗

MHC class I genes in a New World primate, the cotton-top tamarin (Saguinus oedipus), have evolved by an active process of loci turnover.

Lymphocytes of a New World primate, the cotton-top tamarin (Saguinus oedipus), express classical G-related major histocompatibility complex (MHC) class I molecules with unusually limited polymorphism and variability. Three G-related loci, an F locus, an E locus, and two pseudogenes (So-N1 and So-N3) have been identified by cDNA library screening and extensive PCR analysis of both cDNA and genomic DNA from the cotton-top tamarin. Furthermore, each genus of the subfamily Callitrichinae (tamarins and marmosets) appears to express its own unique set of MHC class I genes, likely due to a rapid turnover of loci. The rapid emergence of unique MHC class I genes in the Callitrichinae genera, resulting from an active process of duplication and inactivation of loci, may account for the limited diversity of the MHC class I genes in the cotton-top tamarin. To determine the nature of the entire complement of MHC class I genes in the cotton-top tamarin, we synthesized a genomic DNA library and screened it with MHC class I-specific probes. We isolated nine new MHC class I pseudogenes from this library. These newly isolated tamarin G-related MHC class I pseudogenes are not closely related to any of their functional counterparts in the tamarin, suggesting that they do not share a recent common ancestral gene with the tamarin's currently expressed MHC class I loci. In addition, these tamarin sequences display a high rate of nonsynonymous substitutions in their putative peptide binding region. This indicates that the genes from which they have derived were likely subject to positive selection and, therefore, were once functional. Our data support the notion that an extremely high rate of loci turnover is largely responsible for the limited diversity of the MHC class I genes in the cotton-top tamarin.

3' Untranslated Regions↗

ERIC and REP-PCR banding patterns and sequence analysis of the internal transcribed spacer of rDNA of Stemphylium solani isolates from cotton.

The genetic diversity of the Stemphylium solani isolates from cotton was assessed by Enterobacterial Repetitive Intergenic Consensus (ERIC) and Repetitive Extragenic Palindromes (REP)-PCR fingerprinting. Twenty eight monosporic isolates of S. solani from cotton were used along with five isolates from tomato and one isolate of Alternaria macrospora from cotton for comparison. The dendrogram obtained revealed clear differences between the cotton and tomato isolates as well as between the tomato isolates from different geographic regions. The genetic relationships among S. solani isolates were also analyzed by sequencing the internal transcribed spacer (ITS) region of four isolates representing the three ERIC and REP groups. The tomato isolate from the State of São Paulo showed a distinct ITS sequence from that of the cotton isolates and tomato isolate from the State of Goiás, giving evidence that it belongs to a different genotype of S. solani. This is the first report of the entire sequence of the ITS1-5.8S-ITS2 regions of S. solani.

Ascomycota↗

Longitudinal changes in inflammatory markers in nasal lavage of cotton workers. Relation to endotoxin exposure and lung function changes.

OBJECTIVES: The aim of this study was to evaluate time-related changes in inflammatory markers in nasal lavage (NAL) in relation to endotoxin exposure and lung function changes. METHODS: Endotoxin in cotton dust is well known to elicit proinflammatory mediators involved in acute or chronic respiratory inflammation due to cotton dust. We applied nasal lavage as well as lung function and exposure measurements in a small group (n = 11) of cotton workers during 6 weeks of observation (after 2 weeks free of exposure). Several inflammatory markers in NAL, such as interleukin 8 (IL-8), IL-6, and down-stream mediators, including soluble tumor necrosis factor receptor 75 (sTNF-R75) and soluble intercellular adhesion molecule 1 (ICAM-1), cell counts, marker of epithelial response (uric acid), and marker of plasma exudation (albumin) were measured. RESULTS: At the exposure levels examined in this study [cotton dust, geometric mean (GM) = 1.10 mg/m3; endotoxin, GM = 2,869 EU/m3], several inflammatory markers in NAL such as IL-8, sTNF-R75, and albumin tended to increase through the measurement period with increasing airborne endotoxin concentration (in six workers who had paired measurements in weeks 1 and 6). Although no statistical significance was reached, airborne endotoxin exposure and all inflammatory markers in NAL were relatively higher in cotton workers with a 6-week lung function decrease. CONCLUSION: The data suggest that inflammatory markers in NAL are associated with repeated airborne endotoxin exposure.

Adult↗

Functional characterization of Gossypium hirsutum profilin 1 gene (GhPFN1) in tobacco suspension cells. Characterization of in vivo functions of a cotton profilin gene.

Cotton fiber is an extremely long plant cell. Fiber elongation is a complex process and the genes that are crucial for elongation are largely unknown. We previously cloned a cDNA encoding an isoform of cotton profilin and found that the gene (designated GhPFN1) was preferentially expressed in cotton fibers. In the present study, we have further analyzed the expression pattern of GhPFN1 during fiber development and studied its cellular function using tobacco suspension cells as an experimental system. We report that expression of GhPFN1 is tightly associated with fast elongation of cotton fibers whose growth requires an intact actin cytoskeleton. Overexpression of GhPFN1 in the transgenic tobacco cells was correlated with the formation of elongated cells that contained thicker and longer microfilament cables. Quantitative analyses revealed a 2.5-3.6 fold increase in total profilin levels and a 1.6-2.6 fold increase in the F-actin levels in six independent transgenic lines. In addition to the effect on cell elongation, we also observed delayed cell cycle progression and a slightly lower mitotic index in the transgenic cells. Based on these data, we propose that GhPFN1 may play a critical role in the rapid elongation of cotton fibers by promoting actin polymerization.

Actin Cytoskeleton↗