The in vivo incorporation of radioactivity from [14C]GlcN, [14C]GalN, [14C]Glc and [14C]Gal, for different time intervals between 1 and 240 hr into whole tissues, acetone extracted tissues and MPS-P of the different parts of the reproductive system of the female fowl was studied. The incorporation of radioactivity was much more extensive when [14C]GlcN was injected than when [14C]GalN was injected. The incorporation of radioactivity was much more extensive when [14C]HexN was injected than when the corresponding [14C]Hex was injected. This difference of incorporation was greater in the MPS-P than in the fresh or acetone extracted tissues. A comparison was undertaken in the extent that radioactivity was incorporated among the different parts of the reproductive system of the fowl when [14C]HexN and 14C[Hex] were administered.

The objective of this work was to survey and compare the composition of the parts of the reproductive system of the female fowl in glycosaminoglycans and proteoglycans. Those parts analyzed were ovary, infundibulum, magnum, isthmus, shell gland and vagina. Methods of analysis included cellulose acetate electrophoresis, infrared spectroscopy, colorimetry, amino acid determination and scanning electron microscopy. Concentrations of glycosaminoglycans were higher in vagina, ovary, infundibulum and isthmus than in shell gland and magnum. Glycosaminoglycans may be important in those parts of the reproductive tract which contribute membraneous and mucosal material to the descending egg, and where fertilization of the egg occurs.

Gas liquid chromatography using a methylation and silylation technique is a viable method for the quantitative determination of monosaccharides in serum. Analyses of sera of normal and estrogen-treated birds by this method indicated that estrogen administration increased uronic acid, fucose, galactose, mannose, and sialic acid contents and decreased glucose content of the serum. The greatest of the above increases was that of uronic acid.

Bovine, porcine and avian EMP were isolated and compared for some physical and chemical properties. Some differences in the compositions of three EMPs were observed. The avian EMP contained less carbohydrate than the bovine and porcine EMPs. Some differences in the monosaccharide distributions for the three preparations were revealed. The profiles obtained by SDS-polyacrylamide gel electrophoresis of the preparation indicated a complex (and different for each preparation) nature of the component polypeptides and glycopeptides.

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.

The incorporation of radioactivity from D-[1-14C]glucosamine and D-[1-14C]galactosamine into the tissues of skin, comb, wattle, liver, kidney, spleen, lung, heart, ovaries, egg yolk, infundibulum, magnum, isthmus, shell gland, and vagina of the White Leghorn laying hens was studied for different time intervals up to 120 h. A total of 28 laying hens were involved in this experimentation. The radioactivity was measured in the above whole tissue, the acetone extracts, the acetone-extracted tissues, the mucopolysaccharide-peptide complex (MPS-P), and in individual glycosaminoglycan (GAG) isolated from skin, comb, liver, kidney, and egg yolk. The radioactivities of the expired CO2 of the eggs and of the excreted urine and faeces were also measured in some cases. There was an increase in 14C associated with MPS-P with increasing time of experiment in all the tissues studied except in the case of the liver. The rate of increase of radioactivity was dependent on the tissue. A useful scheme for the study of the speed of the formation of GAG is to consider the acetone extract and the acetone-extracted tissue as macrocompartments from where the GAG draw hexosamines for their formation.

Mucopolysaccharide-peptide complexes (MPS-P) from skin, comb, wattle, liver, kidney, spleen, lung, heart, ovaries, egg yolk, infundibulum, magnum, isthmus, shell gland, and vagina of the White Leghorn laying hens were isolated after extraction with acetone, papain hydrolysis, and cetyl pyridinium chloride precipitation. They were analyzed for characteristic components of the glycosaminoglycan (GAG) and carbohydrate parts of glycoproteins and for amino acids. The infrared spectra of the MPS-P were studied for detection of the GAG present. The MPS-P were submitted to cellulose acetate electrophoresis for detection of GAG and to column chromatography on an anionic resin for quantitative determination of GAG.

Mucopolysaccharide peptide complexes were isolated from avian skin by enzymatic (pepsin–trypsin or papain) hydrolytic procedures each followed by two ways of precipitation: (a) step-wise addition of ethanol or (b) addition of cetyl pyridinium chloride solution and ethanol. Respective precipitates predominantly contained (a) mucopolysaccharide–peptide complex(es) and (b) carbohydrate part of glycoprotein–peptide complex(es). The fractions were analyzed for hexosamine, hexuronic acid and sugars. The results indicated (a) that avian skin contains substantial amounts of both mucopolysaccharides and glycoproteins and (b) that pepsin–trypsin and papain hydrolyses were capable of liberating from avian skin comparable amounts of materials. Gas–liquid chromatography of the fractions indicated that glucosamine, glucuronic acid and galactose are more prevalent than galactosamine, galacturonic acid and mannose.

1.1. Avian plasma was fractionated by sequential ammonium sulfate precipitation, and the carbohydrate composition of the fractions was determined and compared with fractions of porcine and bovine plasmas.2.2. The incorporation of radioactivity of labeled hexosamines into avian serum protein fractions, red blood cells and liver slices was investigated.3.3. Large differences in the degrees and patterns of association of radioactivity were found when labeled glucosamine, galactosamine and mannosamine were injected in fowl, or incubated with avian red cells or liver slices.4.4. The incorporation of labeled hexosamines by red blood cells in vitro may implicate these nucleated cells in glycoprotein synthesis.

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-17β. Most of the changes occurred in the male. Estrogen administration brought statistically significant increases in die 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.

1. 1. Sera of sockeye salmon (Oncorhynchus nerka) were subjected to cellulose acetate and acrylamide gel electrophoresis and the obtained patterns of distribution of proteins and glycoproteins were compared with the corresponding patterns of the bovine serum. 2. 2. It appeared that the distribution of glycoproteins among the proteins of the various electrophoretic zones in the serum of the salmon were more evenly distributed than that in the serum of the domestic cattle (Bos typicus). 3. 3. Plasmas of spring salmon (O. tshawytscha) and cow were fractionated with ammonium sulfate and the fractions were analyzed and chromatographed on carboxymethyl cellulose columns. 4. 4. Differences in the chemical composition of the fractions and of the chromatographic patterns were obvious between salmon's and bovine plasmas.

The mucopolysaccharide–peptide complexes from White Leghorn skin, comb, wattle, heart, liver, and testis were isolated and analyzed for constituent carbohydrate and amino acid units. Individual mucopolysaccharides in each tissue were characterized using several techniques. All tissues examined, except liver, contained at least two mucopolysaccharides, including a low molecular weight component. The low molecular weight component appeared to be the only substance isolated from liver. The incorporation of D-[1- 14 C]hexosamine into acetone-dried tissues and mucopolysaccharide–peptide complexes was investigated. Intravenously injected D-[1- 14 C]galactosamine accumulated in the liver to a much greater extent than D-[1- 14 C]glucosamine. In addition, evidence was provided which indicated that glucosamine was converted to nonhexosamine products. The increase of radioactivity associated with mucopolysaccharide–peptide complexes with increasing time coincided with a decrease in liver radioactivity.

The incorporation of [1-14 C] glucosamine into the serum proteins of White Leghorn was studied following intravenous and intraperitoneal injections. Although the activity curves varied from bird to bird, their maximum occurred approximately two hours after intravenous injection and eight hours after intraperitoneal injection. Electrophoretic distribution of protein, hexosamine and activity were studied following agar electrophoresis. The distributions of hexosamine and activity were not identical. It was suggested that glucosamine has different turn over rates, in different glycoproteins.

The work of Allison and Humphery (1) provided what appears to be the first evidence that the sieving effect of a biogel could be used for the estimation of the molecular weights of polymers. While studying antibody-antigen diffusion coefficients, they noted that, as agar concentration in the gel increased, the size of the antigen which could migrate was decreased, and using this observation they were able to estimate, with a gel of proper agar concentration, the molecular weights of several antigens to within 30%. Smithies (2), using starch gel electrophoresis, suggested that a relationship existed between relative retardation of the mobility of proteins and their molecular size. Both Biogel P300 and DEAE-Sephadex have been used as molecular sieves in the estimation of molecular weights of some acidic polysaccharides by Anderson et al. (3). The most widely used medium for molecular weight estimation of proteins and polypeptides by electrophoresis is the acrylamide gel (4–13). An extensive study on the reliability of this technique has recently been published (14). Following the separation of acidic mucopolysaccharides on polyacrylamide gel (15), the possibility of estimating the molecular weights of acidic mucopolysaccharides from their relative mobilities was investigated.

The levels of hexosamine, sialic acid, fucose, and protein in serum of sockeye salmon (Oncorhynchus nerka) and, to a limited extent, in sera of coho salmon (O. kisutch) and chinook salmon (O. tshawytscha) at two reproductive stages, were determined. Hexosamine, sialic acid, fucose, hexose, seromucoid, and protein content of sexually maturing (early) and mature (spawning) sockeye salmon were studied and a comparison was attempted with the corresponding composition of bovine serum. Content of the above serum constituents was lower in spawning than in maturing populations. Protein content was much less, hexosamine a little less, and sialic acid higher, in the sera of sockeye salmon than in bovine serum. The protein–carbohydrate complex of serum appeared to contain more hexosamine and much more sialic acid than the protein–carbohydrate complex of bovine serum. Furthermore, the sialic acid-to-hexosamine ratio was much higher in sera of salmon than in bovine serum. Some other sex and reproductive stage differences were detected and reported.

The levels of hexosamine, sialic acid, fucose, and protein in serum of sockeye salmon (Oncorhynchus nerka) and, to a limited extent, in sera of coho salmon (O. kisutch) and chinook salmon (O. tshawytscha) at two reproductive stages, were determined. Hexosamine, sialic acid, fucose, hexose, seromucoid, and protein content of sexually maturing (early) and mature (spawning) sockeye salmon were studied and a comparison was attempted with the corresponding composition of bovine serum. Content of the above serum constituents was lower in spawning than in maturing populations. Protein content was much less, hexosamine a little less, and sialic acid higher, in the sera of sockeye salmon than in bovine serum. The protein–carbohydrate complex of serum appeared to contain more hexosamine and much more sialic acid than the protein–carbohydrate complex of bovine serum. Furthermore, the sialic acid-to-hexosamine ratio was much higher in sera of salmon than in bovine serum. Some other sex and reproductive stage differences were detected and reported.

1.1. Species differences were evident in the hexose, hexosamine, sialic acid and fucose contents of bovine, porcine and avian blood sera and in the distributions of proteins and carbohydrates following electrophoreses of these sera in paper, cellulose acetate, polyacrylamide gel and agar.2.2. Stepwise precipitation with ammonium sulfate of porcine plasma and chromatography of the fractions on carboxymethyl cellulose columns yielded products with various carbohydrate compositions some of which have high carbohydrate contents.

Estrone concentrations in whole peripheral blood of seven pregnant cows that calved normally were measured daily from at least three but not more than six days prepartum to four days postpartum. The highest average daily value was 8.2±1.2ng per milliliter at five days prepartum. Blood estrone declined nearly continuously up to one day prepartum, but onset of this decline varied from one to five days before parturition. There was a relatively steep and significant decline between one day prepartum and one day postpartum. The lowest average blood estrone was four days postpartum, beyond which no samples were collected. A heifer that calved abnormally displayed an abnormally high blood estrone at the time of calving.

Blood estrone concentrations of 65 pregnant dairy cows were measured by an Ittrich-Kober fluorometric procedure. Average estrone increased slowly from 1.2ng/ml at 16 to 14 weeks prepartum to 2.5ng/ml at 8 to 6 weeks prepartum, and then more rapidly to 4.8ng/ml at 4 to 2 weeks before parturition. The highest average, 8.3ng/ml, was 5 days preparturn; no sanlples were taken closer to parturition. The lowest average (0.7ng/ml) mas 5 days postpartum.

The compositions of fractions glycosaminoglycuronans and carbohydrate parts of glycoproteins obtained after proteolytic hydrolysis, alcohol precipitation, purification and ethanol fractionation of avian skin, heart, liver and bone, of bovine heart and of ovine heart, lung and liver were determined. Cellulose acetate and acrylamide gel electrophoreses of the fractions indicated that most of the glycosaminoglycuronans present in the fractions had mobilities very near to the mobilities of hyaluronic acid and chondroitin sulfate. a glycosaminoglycuronan present in ovine liver and lung had a mobility equal to heparin, while one from ovine lung migrated faster than heparin. As a result of this and a previous investigation, it is suggested that a gross analytical differentiation, based on proteolytic hydrolyses and on fractional precipitation with ethanol, of the glycosaminoglycuronans and the carbohydrate part of the glycoproteins of the tissues is possible.