Obtaining of a rich-cellulose material from black wattle (Acacia mearnsii De Wild.) bark residues.

Black wattle (Acacia mearnsii De Wild.) barks are residues produced by tannin industries in huge quantities, which are normally discharged on environmental or used for energy production. Therefore, this study aimed to evaluate the use of black wattle bark residues as a raw material on obtaining of a rich-cellulose material by alkaline (MET1), acetosolv (MET2), and organosolv (MET3) procedures. The results obtained indicated that the alkaline methodology, followed by a bleaching step (MET1), promoted klason lignin and hemicellulose removals more efficiently. It was possible to observe that better results were achieved using NaOH concentration of 6% (wt%), at 65 °C for 2.5 h, presenting a yield of 63.24 ± 1.25%, and a reduction on klason lignin content of almost 90.45%. Regarding the bleaching step, it was possible to obtain a material free of non-cellulosic compounds with a yield of 78.28 ± 1.48%. Thermogravimetric analysis indicated the removal of lignin and hemicellulose as well as an increase in cellulose degradation temperature, due to changes in crystalline phases. According to X-ray diffraction (XRD), the procedures employed have led to an increase in crystallinity from 66.27 to 91.78% due to the removal of non-cellulosic compounds. Scanning electron microscopy (SEM) showed morphological alterations in accordance with the removal of non-cellulosic compounds.

Chicken Combs and Wattles as Sources of Bioactive Peptides: Optimization of Hydrolysis, Identification by LC-ESI-MS2 and Bioactivity Assessment.

The production of bioactive peptides from organic by-waste materials is in line with current trends devoted to guaranteeing environmental protection and a circular economy. The objectives of this study were i) to optimize the conditions for obtaining bioactive hydrolysates from chicken combs and wattles using Alcalase, ii) to identify the resulting peptides using LC-ESI-MS2 and iii) to evaluate their chelating and antioxidant activities. The hydrolysate obtained using a ratio of enzyme to substrate of 5% (w/w) and 240 min of hydrolysis showed excellent Fe2+ chelating and antioxidant capacities, reducing Fe3+ and inhibiting 2, 2'-Azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals. The mapping of ion distribution showed that a high degree of hydrolysis led to the production of peptides with m/z ≤ 400, suggesting low mass peptides or peptides with multiple charge precursor ions. The peptides derived from the proteins of cartilage like Collagen alpha-2(I), Collagen alpha-1(I), Collagen alpha-1(III) and elastin contributed to generation of bioactive compounds. Hydrolysates from chicken waste materials could be regarded as candidates to be used as ingredients to design processed foods with functional properties.

Transcriptome analysis of comb and testis from Rose-comb Silky chicken (R1/R1) and Beijing Fatty wild type chicken (r/r).

Rose-comb was one of the chicken comb-variants first used by Bateson and Punnet in 1902 to demonstrate Mendelian inheritance in animals. Rose-comb is a monogenic trait that has been widely described in chickens. It is caused by a large structural rearrangement that leads to mis-expression of transcription factor MNR2 on chromosome 7. Rose-comb has pleiotropic effects in homozygous roosters, which is associated with poor sperm mobility. It was postulated that this is caused by the disruption of the CCDC108 gene located at the distal inversion breakpoint. In this study, we did the transcriptional profiling of combs and testes from Rose-comb Silky (RS) (R1/R1) and Beijing Fatty (BF) wild type chickens (r/r) using RNA-seq. We obtained 68,694,797 unique mapped reads and over 80% of the chicken genes were covered for each sample. In combs, we found that differentially expressed genes (DEGs) were significantly enriched in the retinol metabolism (RPE65, CYP26A1, and CYP26C1) and hedgehog-signaling pathway (PTCH1, GLI1, and HHIP), while genes related to cell differentiation and morphogenesis were down-regulated in R1/R1 chickens, suggesting that the transient expression of MNR2 might affect the expression of these genes and influence the development of comb tissue. For testes, DEGs were significantly enriched in the GO terms of binding activates and mitochondrial oxidation-reduction reactions. Our results suggested that the CCDC108 might be functionally related with mitochondrial oxidation-reduction reactions and caused subfertility of roosters. Compared with the genome average, the degree of expression variations within the inversion region did not show significant differences. However, DEGs near the breakpoints showed greater expression variance. Our results demonstrated that the large-scale rearrangements affected the gene expression only around the breakpoint in this case.

Effect of ultrasonic degradation of hyaluronic acid extracted from rooster comb on antioxidant and antiglycation activities.

CONTENT: Recently, low-molecular-weight hyaluronic acid (LMWHA) has been reported to have novel features, such as free radical scavenging activities, antioxidant activities and dietary supplements. OBJECTIVE: In this study, hyaluronic acid (HA) was extracted from rooster comb and LMWHA was obtained by ultrasonic degradation in order to assess their antioxidant and antiglycation activities. MATERIALS AND METHODS: Molecular weight (Mw) and the content of glucuronic acid (GlcA) were used as the index for comparison of the effect of ultrasonic treatment. The effects on the structure were determined by ultraviolet (UV) spectra and Fourier transform infrared spectra (FTIR). The antioxidant activity was determined by three analytical assays (DPPH, NO and TBARS), and the inhibitory effect against glycated-BSA was also assessed. RESULTS: The GlcA content of HA and LMWHA was estimated at about 48.6% and 47.3%, respectively. The results demonstrate that ultrasonic irradiation decreases the Mw (1090-181 kDa) and intrinsic viscosity (1550-473 mL/g), which indicate the cleavage of the glycosidic bonds. The FTIR and UV spectra did not significantly change before and after degradation. The IC50 value of HA and LWMHA was 1.43, 0.76 and 0.36 mg/mL and 1.20, 0.89 and 0.17 mg/mL toward DPPH, NO and TBARS, respectively. Likewise LMWHA exhibited significant inhibitory effects on the AGEs formation than HA. DISCUSSION AND CONCLUSION: The results demonstrated that the ultrasonic irradiation did not damage and change the chemical structure of HA after degradation; furthermore, decreasing Mw and viscosity of LMWHA after degradation may enhance the antioxidant and antiglycation activity.

Sonic Hedgehog-signalling patterns the developing chicken comb as revealed by exploration of the pea-comb mutation.

The genetic basis and mechanisms behind the morphological variation observed throughout the animal kingdom is still relatively unknown. In the present work we have focused on the establishment of the chicken comb-morphology by exploring the Pea-comb mutant. The wild-type single-comb is reduced in size and distorted in the Pea-comb mutant. Pea-comb is formed by a lateral expansion of the central comb anlage into three ridges and is caused by a mutation in SOX5, which induces ectopic expression of the SOX5 transcription factor in mesenchyme under the developing comb. Analysis of differential gene expression identified decreased Sonic hedgehog (SHH) receptor expression in Pea-comb mesenchyme. By experimentally blocking SHH with cyclopamine, the wild-type single-comb was transformed into a Pea-comb-like phenotype. The results show that the patterning of the chicken comb is under the control of SHH and suggest that ectopic SOX5 expression in the Pea-comb change the response of mesenchyme to SHH signalling with altered comb morphogenesis as a result. A role for the mesenchyme during comb morphogenesis is further supported by the recent finding that another comb-mutant (Rose-comb), is caused by ectopic expression of a transcription factor in comb mesenchyme. The present study does not only give knowledge about how the chicken comb is formed, it also adds to our understanding how mutations or genetic polymorphisms may contribute to inherited variations in the human face.

Competitive protein adsorption on polysaccharide and hyaluronate modified surfaces.

Adsorption of bovine serum albumin (BSA) and fibrinogen (Fg) was measured on six distinct bare and dextran- and hyaluronate-modified silicon surfaces created using two dextran grafting densities and three hyaluronic acid (HA) sodium salts derived from human umbilical cord, rooster comb and Streptococcus zooepidemicus. Film thickness and surface morphology depended on the HA molecular weight and concentration. BSA coverage was enhanced on surfaces in competitive adsorption of BSA:Fg mixtures. Dextranization differentially reduced protein adsorption onto surfaces based on oxidation state. Hyaluronization was demonstrated to provide the greatest resistance to protein coverage, equivalent to that of the most resistant dextranized surface. Resistance to protein adsorption was independent of the type of HA utilized. With changing bulk protein concentration from 20 to 40 µg ml(-1) for each species, Fg coverage on silicon increased by 4x, whereas both BSA and Fg adsorption on dextran and HA were far less dependent on protein bulk concentration.

Copy number variation in intron 1 of SOX5 causes the Pea-comb phenotype in chickens.

Pea-comb is a dominant mutation in chickens that drastically reduces the size of the comb and wattles. It is an adaptive trait in cold climates as it reduces heat loss and makes the chicken less susceptible to frost lesions. Here we report that Pea-comb is caused by a massive amplification of a duplicated sequence located near evolutionary conserved non-coding sequences in intron 1 of the gene encoding the SOX5 transcription factor. This must be the causative mutation since all other polymorphisms associated with the Pea-comb allele were excluded by genetic analysis. SOX5 controls cell fate and differentiation and is essential for skeletal development, chondrocyte differentiation, and extracellular matrix production. Immunostaining in early embryos demonstrated that Pea-comb is associated with ectopic expression of SOX5 in mesenchymal cells located just beneath the surface ectoderm where the comb and wattles will subsequently develop. The results imply that the duplication expansion interferes with the regulation of SOX5 expression during the differentiation of cells crucial for the development of comb and wattles. The study provides novel insight into the nature of mutations that contribute to phenotypic evolution and is the first description of a spontaneous and fully viable mutation in this developmentally important gene.

Effect of estrogen and 2,3,7,8-tetrachlorodibenzo-rho-dioxin (TCDD) on plasma fatty acids of immature male chickens (Gallus domesticus).

This study investigated the effects of 2,3,7,8-tetrachlorodibenzo-rho-dioxin (TCDD) and estrogen on plasma lipids in immature male chickens. Fatty acids were quantified in plasma collected on day 14 from chickens injected with either: Estrogen plus TCDD-1 mg estradiol cypionate /kg body wt. daily for 3 days and 50 microg TCDD/kg body wt. on day 4; Estrogen--1 mg estradiol cypionate/kg body wt. daily for 3 days and vehicle only on day 4; TCDD-vehicle only for 3 days and 50 microg TCDD/kg body wt. on day 4; or Vehicle--same volume of appropriate vehicle for 4 days. TCDD treatment alone increased the plasma concentrations of total triacylglycerides and of the specific fatty acids 14:0, 15:0, 18:0, 18:2n6, 18:3n3, 20:0, 20:1n9, 20:2n6, 20:3n6, 20:5n3 and 22:1n9, compared with vehicle treatment. The concentration of 22:6n3 was increased in all plasma lipid classes of the estrogen group compared with the vehicle group, but was not increased in the estrogen plus TCDD group. Overall, TCDD treatment alone increased plasma lipids, possibly as a result of decreased clearance or utilization; whereas estrogen plus TCDD treatment antagonized estrogen-induced increases in 22:6n3 but did not cause hyperlipidemia.

Immunocytochemical localization of androgen receptor in the comb, uropygial gland, testis, and epididymis in the domestic chicken.

Nuclear, but not cytoplasmic androgen receptors (AR), were localized immunocytochemically in the comb, uropygial (preen) gland, testis, and epididymis of juvenile and adult cockerels. Androgen receptor immunoreactivity (AR-ir) was seen in the comb, in the stratum germinativum of the epidermis and in fibromucoid cells in the dermis in juvenile and adult cockerels. AR-ir was observed in the glandular epithelial (sebum-producing) cells lining the peripheral and middle sections of the tubules in the uropygial gland. AR-ir was not detected in innermost part of the tubules which conduct the sebum to the surface of the skin. AR-ir labeling was not observed in the uropygial gland of juveniles. In the testis, AR-ir was seen in the Leydig cells, in adults but not juveniles. The epithelial cells lining the tubules in the epididymis contained AR-ir in both juveniles and adults.

The effect of furazolidone on fertility of male chickens.

In mature chickens, furazolidone (0.4% w/w, 10 days) decreased the weight of the testes, but did not affect significantly the concentrations of testosterone in testes and plasma, nor the concentrations of ascorbic acid, protein or cholesterol in the testes. Feeding furazolidone at a concentration of 0.08% w/w for 10 days decreased significantly the weights of the testes, wattles and combs. Treatment also produced significant reductions in the concentration of testosterone in plasma and testes, and some reductions in ascorbic acid, protein and cholesterol concentrations in the testes. Administration of furazolidone by crop tube at doses of 40 or 80 mg/kg for five days caused significant decreases in the concentrations of testicular and plasma testosterone, and reductions in ascorbic acid, protein and cholesterol concentrations in the testes. The sizes of the testes, wattles and combs were also significantly reduced. Monoamine oxidase activity in the testes was significantly inhibited by furazolidone. In all the treated birds testicular concentrations of 5-hydroxy-tryptamine were significantly raised, except in those fed the drug at a dose of 0.04% w/w for 10 days.

Studies on the in vitro incorporation of radioactivity from 1-14C-labeled D-hexosamines in the mucopolysaccharide-peptide complexes of avian tissues.

The incorporation of D-[1-14C]-labeled glucosamine (GlcN) and D-[1-14C]-labeled galactosamine (GalN) into mucopolysaccharide-peptide complex(es) (MPS-P) and the rate of 14CO2 production by tissue slices of skin, comb, liver, kidney, shell gland, and magnum from laying hens were studied during a 12 hr period. The D-[1-14C] GlcN was metabolized at a faster rate than D-[1-14C] GalN. No 14CO2 was produced by skin and comb tissues incubated with D-[1-14C]GalN for 12 hr. The amount of 14C associated with the acetone extract of the tissues, acetone-extracted tissues, and MPS-P of the tissue increased with increasing incubation time, but generally the increase was highest in the MPS-P. A comparison among the tissues indicated that the radioactivity present in CO2 and MPS-P was highest in the shell gland and lowest in comb tissue slices. The rates of incorporation of 14C-hexosamine (HexN) into MPS-P by tissue slices appeared to be in general agreement to those of intact animals.

Connective tissue constituents of the fowl. Effects of exogenous estrogen.

The levels of connective tissue constituents (hexosamine, hexuronic acid, hexose and hydroxyproline) and the composition of isolated mucopolysaccharide-peptide complexes were determined in some organs and tissue of male and female fowl treated and not treated with estradiol-17beta. Most of the changes occurred in the male. Estrogen administration brought statistically significant increases in the contents of hexosamine and hexuronic acid in the cartilage of the male and of hexose in the spleen of both males and females. It also increased the hydroxyproline in the testis and in the cartilage of the male. Estrogen decreased the content of hexosamine in the combs of both males and females, and in the wattle of the female, of hexuronic acid, hexose and hydroxyproline in the liver of the male, and hydroxyproline in the breast muscle of the male. Hexosamine, sialic acid, lipid and protein contents of blood sera of estrogenized birds were substantially higher than that of the controls. Gas liquid chromatography of the lipids of the tissues indicated that estrogen administration brought about an increase in the proportion of the unsaturated fraction to the total fatty acid content.

In vivo requirement for silicon in articular cartilage and connective tissue formation in the chick.

Studies were undertaken to determine further effects of silicon deficiency in the chick. The diet and experimental conditions were the same as those used in previous studies to demonstrate the essentiality of silicon for growth and development. Skeletal and other abnormalities involving glycosaminoglycans in formation of articular cartilage and comb connective tissue were found to be associated with silicon deficiency. The bones of 1 day-old deutectomized cockerels fed a silicon supplemented diet and killed at 4 weeks of age had significantly greater amounts of articular cartilage and water as compared with the silicon deficient group and also a greater proportion of hexosamine in the cartilage. The greater water content in bones of the silicon supplemented chicks coincided with a larger content of glycosaminoglycans in the articular cartilage. A similar relationship was obtained in cockerel comb. In addition to larger amounts of connective tissue and of total hexosamine in combs of the supplemented group, a higher percentage of hexosamine and a higher silicon content was found. These findings provide the first evidence for a requirement for silicon in articular cartilage and connective tissue formation and that the site of action of silicon is in the glycosaminoglycan-protein complexes of the ground substance.

Binding of methyltrienolone to various androgen-dependent and androgen-responsive tissues in four animal species.

Comparative sucrose gradient studies of the in vitro binding of dihydrotestosterone (DHT) and of a synthetic androgen, methyltrienolone (R 1881), have been done with the cytosols of various tissues of the rat, mouse, cock and man. With rat prostate cytosol, the amount of R 1881 and DHT binding in the 8-9S region of the gradient was found to be comparable. Specific 8-9S peaks of R 1881 were also found in rat levator ani/bulbocavernosus and skeletal muscles and in the mouse kidney. Only 4-5S peaks could be demonstrated in the cock's comb while DHT under the same conditions showed both 8-9S and 4-5S binding. Binding of R 1881 to the cytosol of the hyperplastic prostate was polydispersed, and showed evidence of the presence of aggregates. Evidence was also found that R 1881 could bind to the progesterone receptor in rat uterus. Our study supports the theory that in a given species the androgen receptors are similar if not identical in all the tissues. The synthetic androgen R 1881 appears to be a useful tool for androgen receptor studies in various animal species provided that the tissue under study contains no progesterone receptor.