Milling Red SWB: A Textile Dye for Staining Epithelial Intercellular Bridges

Fourteen textile dyes were evaluated as histological stains for benign and malignant tissues. An acid dye, Milling red SWB (Acid Red 114, C.I. 23635) was found to possess great affinity for epithelial intercellular bridges. The intercellular bridges were demonstrated clearly when the Milling red SWB was used as a 1% solution in 1% potassium alum after hematoxylin and, also, when methylene blue ZX (Basic Blue 9, C.I. 52015) was used as a counter-stain, without hematoxylin. The intercellular bridges could not be demonstrated in pleomorphic cells which invaded the subepithelial connective tissue.     

A New Basic Triple Salt Containing Magnesium Hydroxide

The quinary system KCl-K₂SO₄-MgCl₂-MgSO₄-Mg(OH)₂-H₂O and associated eight systems K₂SO₄-MgSO₄-Mg(OH)₂-H₂O, MgCl₂-MgSO₄-Mg(OH)₂-H₂O, KCl-MgCl₂-Mg(OH)₂-H₂O, KCl-K₂SO₄-Mg(OH)₂-H₂O, MgSO₄-Mg(OH)₂-H₂O, MgCl₂-Mg(OH)₂-H₂O, K₂SO₄-Mg(OH)₂-H₂O and KCl-Mg(OH)₂-H₂O were investigated at 50° The solid phases of these systems were the new basic triple salt (NS salt B), MgCl₂ · 3Mg(OH)₂ · 8H₂O, MgSO₄ · 5Mg(OH)₂ · 3H₂O, carnallite, leonite, kieserite, hexahydrite, bischofite, potassium chloride, potassium sulfate and magnesium hydroxide and the crystallization fields of these salts in nine systems were determined.     

A New Material Concept for the Red Cell Membrane

The proposition is made that the red cell membrane is a two-dimensional, incompressible material and a general stress-strain law is developed for finite deformations. In the linear form, the character of such a material is analogous to a two-dimensional Mooney material (e.g., rubber), indicating that the molecular structure in the plane of the membrane would consist of long chains, randomly kinked and cross-linked in the natural state. The loose network could be provided by the protein component and the lipid phase could exist interstitially as a liquid bilayer, giving the membrane its two-dimensional incompressibility. The material provides the capability of large deformations exhibited by the discocyte and yet the rigidity associated with the osmotic spherocyte state. It is demonstrated that a membrane of this type can form a sphere at constant area. An illustrative example of the application to single cell discocyte-to-osmotic spherocyte transformations is presented.     

Note on a New Method of Dye Analysis

Positive and negative dye ions may be titrated with one another conductometrically. The method appears to give values reproducible to about one percent. Difficulty has been experienced in preparing a satisfactory primary standard, so that data on the quantitative analysis of stains are not at present available.     

Comparative Spectroscopy of Basic Fuchsin, Schiff Reagent, and a Formalin-Schiff Derivative in Relation to Dye Structure and Staining

Ultraviolet (UV) absorption (200-330 nm) for 0.5 mg/ml aqueous solutions of basic fuchsia unadjusted and those adjusted to pH 0, 1.5, 8.5, and 11 were determined as well as spectra in the visible range (400-675 nm) for solutions with pH 1.9, 2.8, 3.9, 4.7, 5.5, 5.9, 6.5, 7.5, 9.3, 10.4, and 11. The UV absorbance of degassed Schiff reagent containing 1 or 0.5 mg dye/ml, and that of this reagent adjusted to pH 1.5, 2.3, 3.1, 4.5, 6.0, 7.1, and 8.4 were obtained for comparison. The progressive reaction of formalin with degassed Schiff reagent, followed spectrometrically for 2.5 hr, required 2 hr to reach completion. The degassed Schiff reagent contained only traces of-SO₃H as judged from its minimal absorbance between 280 and 295 nm. The UV absorption of this reagent and basic fuchsin in 1 N HCl were found to be identical. The absorbance is that of basic fuchsin reduced by the addition of Cl^- or SO₃H^- to the central methane carbon and H to the amino groups, therefore the leuco structure of basic fuchsin so reduced shows the fomation-NH₃ groups. Infrared (IR) spectra of basic fuchsin, Schiff crystals, and a crystalline formalin-Schiff reaction product support these observations and indicate that the final colored product is a methylsulfonic acid derivative of basic fuchsin. Identical IR spectra were obtained for two types of crystals derived from Schiff reagents indicating that both are the same chemically, although only one became colored on exposure to air. When these crystals were redissolved and SO₂ added, a Schiff reagent of appropriate pH was produced. Since it is derived from a crystalline product, this type of reagent should be useful in histochemical studies     

Neutral Red as a Redox Dye Induces K+ Efflux and Current-Voltage Changes in Eremosphaera,Lemna, and Guard Cells*

Membrane effects of the redox and pH indicator neutral red were studied with the chlorococcal alga Eremosphaera viridis, with Lemna gibba, and with “isolated” guard cells in epidermal peels of Valerianella locusta. Neutral red was extracellularly reduced and caused transmembrane current-voltage changes, an increase in membrane conductance by about 14 nS, an apparent K⁺ net efflux of up to 120 μmol g^?1 FW in 5 min, and an intracellular acidification by up to 0.7 pH units. Neutral red-triggered K⁺ net efflux was most pronounced at low pH, at an E_o more positive than ?200 mV, and without extracellular Ca²⁺. From the experimental data it is concluded that, due to the redox function of the phenazine molecule, extracellular neutral red triggers a trans-plasmalemma e^? transfer, leading to strong membrane depolarization and charge compensating K⁺ net efflux, in addition to some unspecific ion release. As a consequence the intracellular concentration of strong cations relative to strong anions (SID) decreases, resulting in intracellular acidification.     

A Red,Fluorescent Protein from Silkworm

Nondiffusable melanoidin obtained from a glycine-xylose system was heated in aqueous media, and the resulting chemical changes, as affected by the presence of oxygen, pH value, temperature and the addition of transitional metals, were investigated.

Melanoidin, when heated at 90°C in an aqueous solution, caused remarkable discolorization accompanied by the development of fluorescence, oxygen consumption and a noticeable variation of reductone content. Heated melanoidin became polydispersive in molecular weight on gel filtration chromatograms. There appeared reductones, ninhydrin-positive substances, fluorescent substances, aromatic amines, aliphatic carbonyls, and aliphatic primary amines and/or methylene groups in diffusâtes of melanoidin heated in various media.

An increase in pH value favored oxidative discolorization, while an increase in the concentration of transitional metals except Mn²⁺ restrained the discolorization. In the absence of oxygen, heated melanoidin brought about a slight strengthening of color and the accumulation of reductones ca. fifteen times more than the initial level, while in the presence of oxygen the increase of reductone content at the early period was followed by a rapid decrease.

According to the results obtained, the ambivalent reactivity of melanoidin, i.e. polymerization (colorization) and depolymerization (discolorization), was discussed in relation to influencing factors. A mechanism for the production of reductones in heated melanoidin was also proposed.     

Live Cell Multicolor Imaging of Lipid Droplets with a New Dye, LD540

A lipophilic dye based on the Bodipy fluorophore, LD540, was developed for microscopic imaging of lipid droplets. In contrast to previous lipid droplet dyes, it can spectrally be resolved from both green and red fluorophores allowing multicolor imaging in both fixed and living cells. Its improved specificity, brightness and photostability support live cell imaging, which was used to demonstrate by two-color imaging lipid droplet motility along microtubules.     

Staining of Keratin and Keratohyalin with the Reactive Dye Levafix Red Violet E-2BL

Demonstration of keratin in Zenker-fired skin and in tissues stored in formalin can be difficult because such material is unsuitable for histochemical studies. A reactive dye, Levafix red violet E-PBL, proved useful for demonstration of keratohyalin and some types of keratin. Formalin-, Zenker- and methacarn-fired sections were pretreated with alkaline alcohol, stained one hour at 60 C in an aqueous solution containing 0.25% Levafix red violet E-2BL plus 0.25% NaC1, rinsed in buffer solution pH 9, dehydrated and mounted. Keratohyalin granules and stratum corneum were colored red violet; hair and tonofibrils remained unstained. In sections prestained with Mayer's acid hemalum, keratohyalin was dark blue. Sulfonated monoazo dyes without reactive groups colored no tissue structures under the conditions of this technic; apparently, Levafix red violet E-2BL is bound via its reactive group. Polarization microscopic studies suggest binding of Levafix red violet E-2BL by an amorphous matrix of keratin. Correlations with chemical data indicate that the staining patterns parallel the distribution of proteins formed in the stratum granulosum.     

Biodegradation of Azo Dye Methyl Red by Methanogenic Microbial Communities Isolated from Volga River Sediments

Microbiology - Azo dyes are soluble xenobiotics stable under oxidizing conditions, which are widely used in human practice; they are present in liquid and solid industrial and household wastes and...     

A New Form of Myelin Basic Protein Found in Human Brain

Human myelin basic protein was subjected to ion-exchange chromatography at high pH to separate the differently charged components. Polyacrylamide gel electrophoretic patterns of the fractions showed that the less basic fractions 3, 4, and 5 contained significant amounts of a protein somewhat smaller than the more common 18.5-kDa form. Fraction 3 consisted of approximately equal amounts of this smaller polypeptide and component 3, the 18.5-kDa form found in other mammalian myelin basic protein preparations. The two proteins in fraction 3 were separated by fast protein liquid chromatography. Both have blocked N termini and identical C termini (-Met-Ala-Arg-Arg). When the tryptic digests of the two proteins were fractionated by HPLC, the elution profiles were similar, except that four peaks found in the chromatogram of the larger protein were missing from the chromatogram of the smaller one. In addition, an extra peak was found in the elution pattern of the latter chromatogram. Amino acid analysis of the individual tryptic peptides indicated that the smaller protein lacked residues 106-116 (-Gly-Arg-Gly-Leu-Ser-Leu-Ser-Arg-Phe-Ser-Trp-). The deleted portion corresponds exactly to the amino acid sequence encoded by exon 5 of the mouse basic protein gene. This new form of myelin basic protein has a molecular weight of 17,200, calculated from its amino acid composition.     

Contemporary Knowledge of Dye Plant Species and Natural Dye Use in Kurdish Autonomous Region, Iraq

Contemporary knowledge of dye plant species and natural dye use in Kurdish Autonomous Region, Iraq In Kurdistan, natural dyes once played an important role in the life of nomads as they wild-crafted and traded natural dyes for their survival. They learned from their family how to find, harvest, process, and dye with natural dyes. Abandonment of weaving and the nomadic life, and recent changes in the economy have contributed to significant changes in the natural dyeing culture. Traditional knowledge of natural dyeing plants is no longer common among weavers. This study documents the surviving knowledge of dye plant species and assesses the transmission of knowledge between elderly weavers and a younger generation of weavers’ apprentices. Information on dyeing and dyeing plants was elicited through a species recognition task using picture cards, a pile-sorting task, and through in-depth interviews with nomads in the mountains of the Soran district as well as weaving teachers and students in the city of Erbil, Kurdish Autonomous Region, Iraq. Consensus analysis of pile-sorting data supports the hypothesis that informants belong to a single culture. The results confirm the erosion of natural dyeing culture in Kurdistan and stress the need to stimulate knowledge transfer from the elderly, empirical generation to the younger, learning generation. The study also uncovered the existence of a keen interest among the student informants in traditional herbal medicine. If this trend is true for Kurdish urban youth in general, then it could lead to a revival and perpetuation of traditional plant knowledge.     

Paper Chromatography for Analysis Of A Dye Mixture

A paper chromatographic method for the identification and separation of stain components is proposed. Using an alkaline butanol-water solvent and the descending technic, a commercial product, known as “Triosin”, believed to consist of three (undetermined) dyes, has been separated into five components.

Three of the components were apparently parts of a typical eosin Y commercial sample, and the other two were orange G and erythrosin B. By means of elution of the components from the paper, followed by spectrophotometric analysis, it was possible to reconstitute a mixture which appeared to be identical with Triosin in spectrophotometric, chromatographic, and histological properties.     

Experimental Cyanine Red: A New Stain for Nucleic Acids and Acid Mucopolysaccharides

A new cationic dye, experimental cyanine red (du Pont), with an absorption maximum of 536 mμ and a pH of 2.9 in 0.5% aqueous solution, is shown to be suitable for staining nucleic acids and tissue materials presumed to contain acid mucopolysaccharides. Mammalian tissues fixed in 10% neutral buffered formalin or Bouin's fluid are dehydrated, embedded in paraffin, sectioned, mounted, deparaffinized, passed through ethanols to water, and stained for 3-30 min in 0.5% experimental cyanine red in water. Differentiation and dehydration in 3 changes (about 1 min each) of n-butanol is followed by clearing in xylene and mounting in resin.     

New Zealand Lepidoptera: Basic biogeography

Abstract

The composition of the New Zealand Lepidoptera fauna is briefly described, and affinities within Tortricidae analysed. Sixty-five genera are used to indicate four possible sets of relationships, but shortcomings of taxonomic interpretations are outlined. Except for species with a wide dispersal range (some 6% of the fauna) most show affinities consonant with terrane biogeography. The panbiogeographic methodology is a potent tool in systematics, giving direction to the research effort.     

Degradation of Sulphonated Azo Dye Red HE7B by Bacillus sp. and Elucidation of Degradative Pathways

Bacteria capable of degrading the sulfonated azo dye Red HE7B were isolated from textile mill effluent contaminated soil. The most efficient isolate was identified as Bacillus sp. Azo1 and the isolate could successfully decolorize up to 89 % of the dye. The decolorized cultural extract analyzed by HPLC confirmed degradation. Enzymatic analysis showed twofold and fourfold increase in the activity of azoreductase and laccase enzymes, respectively, indicating involvement of both reductive and oxidative enzymes in biodegradation of Red HE7B. Degraded products which were identified by GC/MS analysis included various metabolites like 8-nitroso 1-naphthol, 2-diazonium naphthalene. Mono azo dye intermediate was initially generated from the parent molecule. This mono azo dye was further degraded by the organism, into additional products, depending on the site of cleavage of R–N=N–R molecule. Based on the degradation products identified, three different pathways have been proposed. The mechanism of degradation in two of these pathways is different from that of the previously reported pathway for azo dye degradation. This is the first report of a microbial isolate following multiple pathways for azo dye degradation. Azo dye Red HE7B was observed to be phytotoxic, leading to decrease in root development, shoot length and seedling fresh weight. However, after biotreatment the resulting degradation products were non-phytotoxic.