Mucopolysaccharides Isolated from Human and Cow Colostrums

Mucopolysaccharides were isolated from both human and cow colostrums. Each of the fractionated mucopolysaccharides was considered to be homogeneous from behaviors in chromatography, electrophoresis and sedimentation pattern. The fractions isolated from human colostrum were found to contain 51.0~78.3% carbohydrates consisting of d-galactose, 2-amino-2-deoxy-d-glucose, N-acetylneuraminic acid, l-fucose and d-glucose, and 31.6~11.0% peptides consisting of 16 kinds of amino acids. The sedimentation constants, s_{20, w}, of these fractions were in the range of 0.75 to 1.73 S. The fraction isolated from cow colostrum was found to contain 19.3% carbohydrates consisting of d-galactose, 2-amino-2-deoxy-d-glucose and N-acetylneuraminic acid, and 65.2% peptides or proteins consisting of 18 kinds of amino acids. The sedimentation constant, s_{20, w}, of the fraction was 3.68 S.     

Histochemical Tests for Polyphenols in Plant Tissues

A satisfactory histochemical test for polyphenols in fresh plant tissues is described. The test is based upon a colorimetric method for phenolics using a nitrous acid reaction. Certain catechol derivatives are characterized by the formation of an intense cherry-red in fresh sections of plant tissues with the reagents used. The red color is sufficiently intense to be readily observed microscopically at 100X. Results obtained with other tests also are described.     

Histochemical and Autoradiographic Studies on the Effects of Aging on the Mucopolysaccharides of the Periosteum

An autoradiographic study was made using S³⁵-sulfate for the localization, distribution, and variation in the mucopolysaccharide content of the femoral periosteum of rats from birth to old age. The mucopolysaccharides were also studied histochemically, using toluidine blue O, Rinehart and Abu'l-Haj's colloidal iron method, and the periodic acid-Schiff reaction, before and after hyaluronidase treatment. Autoradiograms revealed the uptake of S³⁵ particularly in the vicinity of the preosseous zone and adjacent osteoblasts. This labelling was highest at the period of rapid bone growth. With increasing age, the S³⁵ uptake became progressively less. The preosseous zone showed γ-metachromatic staining at all ages after treatment with toluidine blue. Active osteoblasts were mostly orthochromatic, however, β-metachromasia was exhibited at a later age. Abundant amounts of intra- and extracellular mucopolysaccharides of both the acid and neutral type were demonstrated in the periosteum. S³⁵ uptake and γ-metachromasia show the presence of sulfated mucopolysaccharides, of which chondroitin sulfate predominates in the preosseous zone. Since S³⁵ uptake is high in active osteoblasts, the inability to demonstrate metachromasia in osteoblasts may indicate either that chondroitin sulfate is liberated as fast as it is being produced, or that it may be present within the cells in a precursor form not detectable by histochemical methods.     

Histochemical Tests for Proteins and Amino Acids; The Characterization of Basic Proteins

For cytophysiological work it is important to have ways of demonstrating proteins and amino acids and especially of characterizing basic and non-basic proteins. The author presents a review of the more usually employed histochemical reactions for amino acids and proteic compounds in general, with several modifications which increase their sensitivity, or specificity and localization. The author describes the histochemical arginine reaction, recently introduced by him, by means of which the characterization of basic and non-basic proteins can be easily accomplished in every laboratory without costly apparatus; this reaction serves also for the demonstration of proteins in general. The application of protein histochemical tests for quantitative purposes is discussed in connection with the characterization of the basic proteins and the determination of the relative concentration and the active metabolic changes of proteic compounds.     

The Effect of Formaldehyde on Tissue Lipids and on Histochemical Reactions for Carbonyl Groups

Histcchemical and chemical evidence indicates that formaldehyde combines with unsaturated lipids at the double bond. The resulting complex contains a free carbonyl group which probably originates from the formaldehyde. The reaction occurs over a wide pH range, and takes place in the absence of oxygen or moisture. The reaction product is visualized by the Schiff reagent, and by the Ashbel-Seligman procedure. In the plasmal procedure, when performed on formalin-treated material, the reaction has the same significance as the pseudo-plasmal reaction, i.e. it denotes the presence of double bonds. The Ashbel-Seligman technic seems to be more sensitive to this complex than the Schiff reagent and shows it more markedly than it does the true plasmals and the atmospherically oxidized unsaturated compounds.     

The Effect of Formaldehyde on Tissue Lipids and on Histochemical Reactions for Carbonyl Groups

Histcchemical and chemical evidence indicates that formaldehyde combines with unsaturated lipids at the double bond. The resulting complex contains a free carbonyl group which probably originates from the formaldehyde. The reaction occurs over a wide pH range, and takes place in the absence of oxygen or moisture. The reaction product is visualized by the Schiff reagent, and by the Ashbel-Seligman procedure. In the plasmal procedure, when performed on formalin-treated material, the reaction has the same significance as the pseudo-plasmal reaction, i.e. it denotes the presence of double bonds. The Ashbel-Seligman technic seems to be more sensitive to this complex than the Schiff reagent and shows it more markedly than it does the true plasmals and the atmospherically oxidized unsaturated compounds.     

Histochemical observations on the female reproductive tract of bats. IV. Mucopolysaccharides in the uterus of three species of bats

The three species of bats studied show glycogen, neutral, sialo- and sulfomucins in the cells of the uterine epithelium and glands. Elaboration of these mucins occurs during anestrus in Pteropus giganteus and during proestrus in Taphozous longimanus. In Megaderma lyra lyra only glycogen is observed during anestrus and proestrus. Neutral, and sialomucins in addition to glycogen are observed during estrus. The concentration of these mucins increases progressively and is maximum at the time of implantation, decreasing thereafter in all the three species of bats. The elaboration and increase of mucins is probably under the influence of hormones of the ipsilateral ovary.     

Some Reactions of Isolated Corn Mitochondria Influenced by Juglone

The effects of juglone on the uptake of O₂ by excised corn roots (Zea mays L., Wf9 cms- T × M14) and isolated corn mitochondria arc reported. The O₂ uptake by excised corn roots, as measured by an O₂ electrode, was inhibited more than 90% after a one-hour treatment of 500 μM juglone. Lesser inhibitions were observed with 50 μM and 250 μM juglone. In a KC1 reaction medium in the absence of inorganic phosphate (Pi), juglone stimulated the rate of O₂ uptake by isolated mitochondria oxidizing NADH, succinate, or malate + pyruvate. In the presence of Pi, juglone concentrations of 3 μM and greater inhibited the state 3 oxidation rates of succinate and malate + pyruvate, lowered respiratory control and ADP/O ratios obtained from the oxidation of NADH, malate + pyruvate, or succinate, and reduced the coupled deposition of calcium phosphate within isolated mitochondria driven, by the oxidation of malate + pyruvate. The inhibition of state 3 O₂ uptake by isolated mitochondria, an oxidative state in which electron transfer is coupled to ATP production, is seen to correlate with the inhibition affected by juglone when applied to tissues in vivo.     

Exact Power of Conditional Tests for Several Binomial Populations

An efficient recursive polynomial multiplication method is proposed for exact unconditional power calculation for unordered 2 × K contingency table with up to moderate sample size. Our method can be applied to the family of cell-additive statistics which includes the Freeman-Halton statistic, the Pearson χ² statistic and the likelihood ratio statistic. We illustrate our proposed method by several numerical examples.     

The Histochemical Localization of Several Enzymes of Soybeans Infected with the Root-knot Nematode Meloidogyne incognita acrita

The sites of activity of alkaline phosphatase, acid phosphatase, esterase, peroxidase, adenosine triphosphatase, and cytochrome oxidase were demonstrated histochemically in fresh sections of ''Lee'' soybeans infected by the root-knot nematode Meloidogyne incognita acrita. Each of the six enzymes was more active at the sites of infection than in adjacent non-infected tissue. During the early stages of infection, an increase in enzyme activity was observed in several cells in the proximity of the lip region of the nematode. However, when definite syncytia were observed, increased enzyme activity was confined primarily within the limits of the syncytium. Increased activity paralleled syncytial development and nematode maturation.     

Histochemical Localization of Several Enzymes in Early Young Embryo of Pinus tabulaeformis

Histochemical localization of adenosine triphosphatase, acid phosphatase and peroxidase in young embryo of Pinus tabulaeformis has been studied. All cells of the embryo proper showed very intense lead phosphate disposite with ATP in the Wachs- tein Meisel medium and with B-glyceropbosphate in Gomori medium. In the suspensor 1–3 suspensor cells closed to embryo proper appear blackening with both the ATP and the B-glycerophosphate, and other cells of the suspensor showed less adenosine triphosphatase and B-glyeerophosphatase. In the young embryo the peroxidase distribution may vary with the developmental stages of the young embryo, for instance cone-shaped embryo proper showed dark blue, and which peroxidase occupies only 1–2 suspensor cells closed to embryo proper. When the young embryo developed further into columnus-shape, the peroxidase was mainly distributed in regions of both the posterior part of embryo proper and just derived suspensor cells. However the anterior part of embryo proper showed slight peroxidase. The experimental results indicate that all of these enzymes mentioned above are mainly present in both the embryo proper and the connecting end of the suspensor, and that these regions show great matabolic activity. From the above mentioned observation, the possible relationship between the young embryo and near surrounding tissue during the development of embryo is discussed.     

Histochemical studies on the mucopolysaccharides of the developing chick kidney. II. Mucopolysaccharides of the collecting tubules and excretory ducts.

Mucopolysaccharides of the collecting tubules and excretory ducts of developing chick kidney were studied histochemically to elucidate the relations, between the changing chemical properties of these compounds and excretory processes during pronephric, mesonephric and metanephric developmental stages. At the pronephric ammonotelic stage the collecting tubules contain only neutral mucopolysaccharides, whereas at the mesonephric ureotelic stage neutral mucopoly saccharides are found till the 9th day. Sialic acids make their appearance in luminal border regions of the mesonephric collecting tubules from the 9th day on, their concentration being highest on the 15th day. Hyaluronic acid is observed from the 16th day on. Its concentration is predominant in hatched young birds and increases with age. The physiological significance of alterations in the mucopolysaccharides contents is discussed.     

Generalized Maximum Range Tests for Pairwise Comparisons of Several Populations

A multiple comparison procedure (MCP) is proposed for the comparison of all pairs of several independent samples. This MCP is essentially the closed procedure with union-intersection tests based on given single tests Q_ij for the minimal hypotheses H_ij. In such cases where the α-levels of the nominal tests associated with the MCP can be exhausted, this MCP has a uniformly higher all pair power than any refined Bonferroni test using the same Q_ij. Two different general algorithms are described in section 3. A probability inequality for ranges of i.i.d. random variables which is useful for some algorithms is proved in section 4. Section 5 contains the application to independent normally distributed estimates and section 6 the comparisons of polynomial distributions by multivariate ranges. Further applications are possible. Tables of the 0.05-bounds for the tests of section 5 and 6 are enclosed.