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A continuum mechanics approach to determining the cellular velocity field with a wool follicle.

A model, based on the principles of continuum mechanics, is presented for the analysis of cell-velocity fields within wool follicles. The model requires specification of three follicle characteristics in the form of spatially varying fields: viscosity, cell density and cell production rate. The viscosity is introduced as an attempt to model both complex intercellular interactions and individual cell deformation as the cells move. It is demonstrated that the distribution of cell production is more important than axial variation in viscosity in determining the overall flow pattern.

Animals↗

Lysinoalanine formation in wool after treatments with some phosphate salts.

Treatment of wool with solutions of sodium tripolyphosphate, potassium tetrapyrophosphate, and sodium phosphate results in the formation of lysinoalanine, the amount of which increases from the first to the last reagent. As this may be due to the different pH, solutions of the three salts of the same pH were also tried. This series of experiments suggests that although the hydroxyl ions contribute to the formation of lysinoalanine, the governing factor is the kind of the anions concerned, as the sodium phosphate acts in a more rigorous may than the two other salts, in spite of the smaller concentration and the faster reduction of the pH of the solution during treatment.

Alanine↗

Effects of maternal nutrition on the initiation of secondary wool follicles in foetal sheep.

The initiation of secondary (S) wool follicles usually takes place between about 95 and 135 days of gestation. Severe underfeeding during the first half of this period did not significantly inhibit the initiation of S follicles, but severe underfeeding during the latter half of this period resulted in a significantly lower number of S follicles and this number was not increased by refeeding ewes to a high level between 132 days and term. A similar reduction in S follicle initiation was effected when ewes were moderately underfed throughout most of pregnancy, but when the ewes were refed between 120 days of gestation and term the numbers of S follicles initiated were not significantly different from those of a group of well-fed animals. It is concluded that S follicle initiation is most affected by maternal undernutrition between about 115 and 135 days of gestation.

Animals↗

Aggregation of wool keratin intermediate filament proteins.

The wool keratin intermediate filament proteins were isolated as their S-carboxymethyl derivatives (S-carboxymethylkerateine A, SCMKA) and purified by gel filtration to remove residual non-helical protein of low molecular weight. The alpha-helix content of purified SCMKA was approximately 62% in agreement with that predicted for the alpha-helical coiled-coil segments from the amino acid sequences of the subunits. In aqueous buffer at pH 11 or in n-propanol (20% v/v) at pH 9.2 very large aggregates are dissociated and SCMKA exists largely as a mixture of the dimer (two-chain coiled-coil of Mr approximately 103,000) and the tetramer. The protein species are not in rapidly reversible equilibrium as judged from gel filtration and sedimentation equilibrium. It is probable that species with a range of association constants are present. The equilibrium is shifted towards the dimer with change of pH from 9.2 to 11 or by the addition of 20% (v/v) n-propanol. The tetrameric proteolytic digestion product which is derived from the 1B segment of the alpha-helical rod section of the keratin molecule dissociates in a similar way to intact SCMKA with increase of pH and in the presence of n-propanol. This indicates the importance of this region of the rod domain in the initial stages of the assembly of the filament. Electrostatic and hydrophobic interactions are implicated in the association of the two-chain coiled-coil to the tetramer both in intact SCMKA and the 1B segment tetramer. The results are discussed in relation to the intact dimeric and tetrameric complexes obtained from other intermediate filament types.

Amino Acids↗

NMR evidence of formation of cyclocystine loops in peptide models of the high sulfur proteins from wool.

We report that in peptide models of the high sulfur proteins of the matrix from wool a disulfide bond forms between two sequential Cys-residues as a result of a simple oxidation procedure. This tiny cyclocystine loop manifests itself in many ways in 1H NMR spectra. The formation of the loop is accompanied, as expected, by conversion of the Cys-Cys peptide bond from its usual trans-configuration into the energetically less favorable cis-configuration. Possible consequences of the formation of cyclocystine loops for the elasticity of the network of the matrix are discussed.

Animals↗

An evaluation of serum sources used in the separation of T- and B-lymphocytes by nylon-wool columns.

Gamma-globulin-free (GG-) newborn-calf, mouse and horse, as well as newborn-calf and calf serums were evaluated as substitutes for fetal bovine serum in the classical nylon-wool procedure used for the separation of T- and B-lymphocytes. The efficacy of the cell separation was evaluated by non-specific esterase staining and cytotoxicity tests for cell surface Thy 1.2 antigen and surface immunoglobulins. The results of these studies indicate that GG- newborn-calf, mouse and horse but not newborn-calf or calf serum could be substituted for fetal bovine serum. The results of these studies also indicate that the ratio of adherent to non-adherent cells is a useful indicator of the efficacy of the cell separation. A ratio of non-adherent to adherent less than 1.0 is indicative of a successful separation.

Animals↗

Growth factor expression in skin during wool follicle development.

A variety of growth factors are likely to be involved in initiation and morphogenesis of wool follicles. To enable direct comparisons of the expression of different growth factors, reverse transcriptase-polymerase chain reactions (RT-PCR) were developed for ovine and murine TGF alpha, TGF beta 1, TGF beta 2, TGF beta 3, IGF1, IGF2, and FGF-2, which could all be carried out on a single cDNA sample. These RT-PCR were used with 16 sheep RNA samples from different foetal stages, neonatal sheep and mouse skin. The mRNAs for these growth factors were detected throughout gestation in sheep skin, except for TGF beta 1 mRNA which was not expressed in 51-day-old skin, but was expressed in 54-day and older samples. Since the first microscopically visible changes of follicle initiation occur around 62 days gestation, these results suggest that TGF beta 1 expression may be a signal for follicle initiation.

Animals↗

Characterization and distribution of epidermal growth factor receptors in the skin and wool follicles of the sheep fetus during development.

We have determined the binding affinity and capacity and relative distribution of epidermal growth factor (EGF) receptors in the skin of the Merino sheep fetus before and during the development of the wool follicle population. Autoradiography of tissue sections incubated with [125I]EGF revealed that label was confined predominantly to the epidermis and dermoepidermal junction before follicle formation, at 30 and 55 d of gestation. During follicle initiation (Days 60 to 65), receptor activity was distributed over the epidermis, including the epidermal aggregations of primordia at the dermoepidermal junction. However, receptor concentrations, as revealed by grain counts of autoradiographs, were reduced in these regions when compared with 55-d skin. The receptor distribution over the epidermis and its derivatives did not alter during subsequent follicle development, although the intensity of labeling increased as the follicles matured. Specific receptor binding was not observed above background levels in the dermis and dermal papillae during all stages of follicle development. At follicle maturation, EGF receptors were widely distributed over the cells of the epidermis and the epidermal derivatives of the cutaneous appendages but were particularly localized in the sebaceous glands and outer root sheath (see also Wynn et al. 1989). EGF immunoreactive material has also been found at these sites (du Cros et al. 1992), suggesting an autocrine role for EGF in the regulation of cell function. It is likely that the differentiation-promoting activities of EGF may predominate over those of growth, because the receptor-bearing cells were not members of rapidly proliferating populations.

Animals↗

Flow behavior of regenerated wool-keratin proteins in different mediums.

Keratin is abundantly present in nature and the major component of hair, wool, feather, nail and horns. Dissolution of keratin is often required when non-textile applications are demanded. However, the low solubility of keratin in water is the major problem. It becomes unstable and precipitated when stored for a long time. Therefore, it is necessary to find a good solvent that provides high stability and easy processibility. In this research, we used formic acid and dimethylformamide (DMF) to dissolve regenerated keratin protein films. It is shown that formic acid is a good solvent for regenerated keratin proteins for the purpose of storage. Transparent and stable regenerated keratin solution is obtained in formic acid.

Animals↗

Integrating transcriptomics and metabolomics reveals the molecular landscape of sperm maturation driven by regional differentiation in the epididymis of Guizhou-Guiqian semi-fine wool sheep.

Epididymal regionalized differentiation is crucial for sperm maturation. However, little is known about the synergistic remodeling mechanisms of different epididymal segments at the transcriptional and metabolic levels during sexual maturation in ruminants (especially sheep). We investigated the caput, corpus, and cauda epididymidis of pre-pubertal (2-month-old) and post-pubertal (7-month-old) Guizhou-Guiqian semi-fine wool sheep using histology, RNA sequencing, and metabolomics. Post-pubertal tissues exhibited increased luminal diameters, cilia lengths, and abundant cauda spermatozoa. Transcriptomic analysis revealed increasing differentially expressed genes (DEGs) along the caput-corpus-cauda axis (4642, 6103, and 7698 DEGs, respectively). Metabolomics detected 786 unique differentially accumulated metabolites (DAMs). Region-specific analysis showed that in the caput, up-regulated pathways (fructose/mannose metabolism; HK2, ALDOA, HKDC1) provide energy and substrates for initial sperm motility. In the corpus, down-regulated genes associated with extracellular matrix and tight junctions suggested epithelial barrier remodeling to establish an immune-tolerant microenvironment. The cauda specifically up-regulated the pentose phosphate pathway (FBP1, GPI) and glutathione metabolism, maintaining redox homeostasis for long-term sperm storage. Additionally, glycerophospholipid metabolism was enriched across all segments, where PEMT, AGPAT5, and LCAT likely regulate sperm plasma membrane fluidity. In conclusion, during sexual maturation, the caput drives energy metabolism and glycosylation, the corpus establishes immune tolerance, and the cauda maintains antioxidant homeostasis. The glycerophospholipid network throughout the across all epididymal segments synergistically remodels sperm membrane. This study reveals the underlying multi-omics regulatory mechanisms of epididymal functional differentiation, providing a theoretical basis for elucidating the molecular mechanisms of sperm maturation in this breed and for the molecular breeding of early reproductive performance in rams.

Animals↗

Determination of peptides and amino acids from wool and beer with sensitive fluorescent reagent 2-(9-carbazole)-ethyl chloroformate by reverse phase high-performance liquid chromotography and liquid chromotography mass spectrometry.

A new method for the sensitive determination of amino acids and peptides using the tagging reagent 2-(9-carbazole)-ethyl chloroformate (CEOC) with fluorescence (FL) detection has been developed. Identification of derivatives was carried out by liquid chromotography mass spectrometry. The chromophore in the 2-(9-fluorenyl)-ethyl chloroformate (FMOC) reagent was replaced by carbazole, which resulted in a sensitive fluorescence lerivatizing agent CEOC. CEOC can easily and quickly label peptides and amino acids. Derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography. Studies on derivatization demonstrate excellent derivative yields over the pH range 8.8-10.0. Maximal yields close to 100% are observed with three- to fourfold molar reagent excess. Derivatives exhibit strong fluorescence and allow direct injection of the reaction mixture with no significant disturbance from the major fluorescent reagent degradation by-products, such as 2-(9-carbazole)-ethanol and bis-(2-(9-carbazole)-ethyl) carbonate. In addition, the detection responses for CEOC derivatives are compared to those obtained with FMOC. The ratios AC(CEOC)/AC(FMOC) = 1.00-1.82 for fluorescence (FL) response and AC'(CEOC)/AC'(FMOC) = 1.00-1.21 for ultraviolet (UV) response are observed (here, AC and AC' are, respectively, FL and UV response). Separation of the derivatized peptides and amino acids has been optimized on a Hypersil BDS C18 column. Excellent linear responses are observed. This method was used successfully to analyze protein hydrolysates from wool and from direct-derivatized beer.

Amino Acids↗

Long-term effects of short-term provision of protein-enriched diets on resistance to nematode infection, and live-weight gain and wool growth in sheep.

Weaner sheep that had been hand-fed on diets containing increasing concentrations of protein for a 9-week period (when uninfected, or infected with Haemonchus contortus) were studied during the next 69 weeks when put on to pasture as a single, unsupplemented flock. During the 9-week period, groups of 12 sheep (six infected, six uninfected) were offered one of five iso-energetic (9.0 MJ kg(-1)) diets containing 10, 13, 16, 19 or 22% crude protein. All sheep were treated with anthelmintic at the end of the 9 weeks and then put out to pasture for 69 weeks, where they were all subject to the same environmental variables including nematode larval challenge. During the grazing period, animals that had previously received the higher protein diets consistently had higher live-weight gain and wool production, higher antibody responses to both H. contortus and Trichostrongylus colubriformis antigenic challenge in vitro, and lower faecal nematode egg counts than did the lambs previously offered the lower protein diets. Faecal egg counts of the grazing sheep that had been artificially infected with H. contortus while being hand-fed were similar to those of the uninfected sheep and there was no interaction between previous infection and dietary protein concentration. We conclude that short periods of enhanced post-weaning nutrition can have long-term and perhaps life-long effects on production.

Animals↗

A critical review of the structural mechanics of wool and hair fibres.

The potential of genetic engineering gives current importance to the need to clarify the relations between structure, properties and performance of wool. There are at present three different models to explain the tensile stress-strain properties of alpha-keratin fibres: a development by Wortmann and Zahn (W/Z) of Feughelman's series-zone model based on the structure of intermediate filaments and a gel-->sol matrix; a new model by Feughelman (F94) based on a matrix of protein globules surrounded by water; and Chapman's model based on the composite mechanics of microfibrils, characterised by critical and equilibrium stresses for the alpha<-->beta transition, in an elastomeric matrix. This paper examines the arguments related to these theories, with additional attention to the Chapman/Hearle (C/H) treatment. The main area of uncertainty, on which more information is much needed, concerns the chemical and physical structure and properties of the matrix, which is composed of keratin-associated proteins. The conclusion is that the C/H model, which is based on reasonable values of input parameters and gives good agreement with a wide range of experimental results, is most likely to be valid, though it needs some additional refinement and incorporation in a total model, which includes larger-scale structural features.

Animals↗

The role of chitosan in wool finishing.

The application of the biopolymer chitosan as an alternative to the conventional contaminating processes in textiles was studied. As chitosan is produced by biomass, it is biodegradable and bioadsorbable. These properties are increasingly important given the current environmental legislation. The main aim of chitosan treatment of oxidised wool fabrics is to improve felting properties and dyeing behaviour.

Animals↗

A study of the influence of structure on the effectiveness of chitosan as an anti-felting treatment for wool.

The effect of chitosan on the resistance of wool fabric to felting on washing has been studied using nine structurally different samples of chitosan. Structural differences examined include molecular weight, level of N-acetylation, and the nature and concentration of homologous N-acyl groups. No strong dependency of shrinkage on molecular weight or level of N-acetylation has been found, but increasing the hydrophobic character of chitosan through the incorporation of a number of long-chain N-acyl groups gives improved anti-felting behaviour, compared to chitosan itself, at the same level of add-on.

Acetylation↗

Gene expression in sheep skin and wool (hair).

We sequenced 2939 ESTs from fetal and adult sheep skin. Stages of gestation were picked to coincide with the major events in skin appendage (wool follicle) formation. Clustering analysis generated a nonredundant set of ESTs 2435 strong (83% nonredundant). Approximately 24% of these gave no hit to NCBI build 29 of the human genome, while 35% were tentatively classified by putative function based on BLASTX hits with a p(N) of <10(-4). In addition to bioinformatics analysis of our ESTs and gene mapping, we have generated a large EST spatial expression data set using in situ hybridization. One thousand one hundred forty-two ESTs have been used for in situ localization; about 31% are from adult sheep skin, 39% from late gestation fetal sheep skin, and 30% from midgestation fetal sheep skin. These probes have been used in over 3000 hybridization experiments. In this report, we summarize the results of in situs on adult sheep skin.

Animals↗

Convenient and rapid detection of pesticides in extracts of sheep wool.

A relatively simple, rapid extraction technique based on acetonitrile was combined with the use of screen-printed electrodes bearing cholinesterases to detect organo-phosphate pesticides from an otherwise intractable matrix, sheep wool. It proved possible to separate, for convenience, the exposure of the electrodes from measurement of their (inhibited) activity. The electrodes were used once and then discarded. Estimation of the extent of inhibition is dependent on reference to the activity of control electrodes. The presence of pesticides in the extracts could be detected with any of three commonly available cholinesterases but the most sensitive enzyme was butyryl cholinesterase from horse serum.

Acetonitriles↗

A further study on the dietary-regulated biosynthesis of high-sulphur wool proteins.

When the diet of sheep is supplemented by the infusion of sulphur-containing amino acids or casein into the abomasum, the newly synthesized wool shows characteristic changes in its amino acid composition, with significant increases in cystine, proline and serine and decreases in aspartic acid and phenylalanine. This modification seems to be due entirely to an alteration in the overall composition of the high-sulphur proteins and to an increase in their proportion in the fibre. These variations are not the result of a change in the composition of individual proteins, but are due to alterations in their relative proportions and to the initiation of the synthesis of ;new' proteins, many of which are extremely rich in cystine. It is suggested that the heterogeneity of the high-sulphur proteins may be due, in part, to similar changes in composition caused by natural variations in the nutrition of sheep.

Abomasum↗