Nile Blue Sulfate Research Articles

History of the hypoblast in salmo.

AbstractSpots printed on the blastodisc of Salmo gairdneri with Nile blue sulfate prior to and during the formation of the germ ring and embryonic shield show during the morphogenetic movements that not many of the germ ring cells are derived from the center of the blastodisc. Evidence suggests that only a few of the loose round cells of the deep part of the ring and of the axial nubbin are produced by local dropout of cells at the margin of the blastodisc, but that most of them arrive there by outward migration from the outer two‐thirds of the disc. The embryonic shield is produced by a convergence of internal cells that starts early in the fifth day at 10° C, augmented by a massive descent of consolidated internal cells from the central one‐third of the blastodisc which then form the forebrain and optic vesicles. The cellular envelope is not involved in an invagination, nor does it take part in the general convergence movements.Chalk grains incorporated into the lower surface of the germ ring and the embryonic shield reveal the destination of the cells which carry them along. The hypoblast of the ring continues to converge toward the shield until the closure of the yolk plug on the ninth day. The epiblast of the ring meanwhile forms the cell layers of the yolk sac. Head mesoderm, pharynx endoderm and notochord materials are assembled directly into the embryonic shield while the germ ring is forming. Additional evidence is presented that these parts are not formed by invagination. Materials of the hypoblast originally placed as far as 90° out on the ring from the early shield reach the trunk endoderm, and materials from the hypoblast of the entire ring are passed along toward and into the somite and lateral plate strips. Their oblique paths of migration, combining both epiboly and convergence, are at marked angles to the simple epibolic paths of the epiblast cells, and it is suggested that the split between epiblast and hypoblast in the germ ring is a result of this shearing effect.

VITAL DYES AS MARKERS FOR BEHAVIORAL AND POPULATION STUDIES OF THE LARCH SAWFLY, PRISTIPHORA ERICHSONII (HYMENOPTERA: TENTHREDINIDAE)

AbstractLarvae of the larch sawfly, Pristiphora erichsonii (Htg.), were fed foliage of tamarack, Larix laricina (Du Roi) K. Koch, treated with solutions of the vital dyes, Nile blue sulphate, rhodamine B, and coriphosphine O, to determine their suitability as markers. Coriphosphine O was highly toxic at the concentrations administered but successful tagging was obtained with the other two dyes. Optimal dye retention and survival were obtained when larvae were fed for 3 days early in the last stadium using Nile blue sulphate and throughout the last stadium using rhodamine B.Both dyes persisted throughout development and were incorporated in the cocoons and the oviposited eggs. They could be readily seen in all stages except for the cocoons and for eggs that contained rhodamine B. The dyes could be detected in cocoons by treating them with cold KOH followed by ethanol. The presence of rhodamine B in the eggs could be ascertained by its yellow fluorescence in ultraviolet.The dyes incorporated in the cocoons are relatively stable and laboratory tests indicate they probably can withstand weathering under normal field conditions over a 1-year period.The use of these dyes for internal marking should prove useful in studying adult dispersal and flight range and as a technique for use in population studies of the larch sawfly.

The use of Nile blue sulphate in the histochemical identification of phospholipids.

Representative extracted lipids were examined visually and spectrophoto-metrically following staining under varying conditions with the oxazine dye Nile blue sulphate. Optimum results in staining both extracted and tissue lipids were obtained with 1% acid-hydrolysed Nile blue prepared by boiling 200 ml of dye with 10 ml of 1% H2SO4 for 4 hr with refluxing. Staining was carried out at 70 C for 30 min. Under these staining conditions, extracted phospholipids were coloured intensely blue by the oxazine base of the dye, except for a synthetic cephalin which behaved as a neutral lipid. Neutral lipids (trigly-cerides and cholesteryl esters) were stained red by the oxazone derivative. Free fatty acids were coloured magneta-red due to a blend of red (oxazone) and blue (oxazine) staining. The procedure recommended clearly distinguished phospholipids from the other main lipid classes in tissue sections. Localisation of phospholipid was improved by also staining sections from which hydrophobic lipids had been remove...

Chromatographic separation and photometric analysis of the components of Nile blue sulphate.

Samples of Nile blue sulphate (Geigy and G. T. Gurr) were separated by silicic acid thin layer chromatography. The blue sulphated oxazine base was found to consist of 3 components with identical absorption spectra in the visible range. These probably represent isomeric forms of the dye. The red oxazone travelled as a single component. At least 6 minor constituents, present in small amount, were identified. Spectrophotometric examination showed that the sulphated oxazine base has 3 absorption maxima. The absorption spectrum is influenced by pH, dye concentration, dye solvent, and physical state. The absorption spectra of the oxazone and free oxazine base were also measured. These showed marked solvatochromic effects, absorption maxima moving to shorter wavelengths with decreasing solvent polarity.

Calcification and its mechanism in the shore-crab, Carcinus maenas (L.)

Crustacean cuticle is strengthened by the deposition of lime. Calcification in general is usually attributed to the combined action of enzymes and to an organic seeding mechanism. The production of alkalinity by electrode action, resulting from diffusion of salt water through a protein membrane containing semiconducting substances, suggests that lime may be precipitated in this way. This theory is examined with particular reference to the shore-crab, Carcinus maenas (L.). Semiconducting materials in protein or cuticle were detected mainly by the conversion of the dye Nile blue sulphate to the alkaline form, the red base, on outward diffusion of salt. This is termed the ‘Nile blue brine’ reaction. When this treatment was applied to protein to which hydroquinone and calcium carbonate had been added, alkalinity was greatly enhanced by the further addition of the bicarbonate or ferrous ion. The reaction is then comparable to that in normal crab cuticle. Calcification of the cuticle in relation to semiconducting material has been studied through the moult cycle. Calcification at each stage occurs in regions which according to the electrochemical theory proposed are normally cathodic, supporting the view that such calcification is electrochemical in origin. The conduction mechanism is discussed with reference to the mechanism of charge transport and polarization. It is concluded that the essentials for calcification of the surface of an organism such as the crab are: processes which maintain the inside electronegative to the outside, the presence of semiconducting complexes in, the cuticle and the presence of sufficient lime in the environment.

Studies on Echinostomatidae (Trematoda) in Malaya. XIV. Body Gland Cells in Cercariae of Echinostoma audyi Lie and Umathevy, and E. lindoense Sandground and Bonne

Cercariae of Echinostoma audyi and E. lindoense have, in addition to cystogenous and penetration gland cells, other gland cells that open on the body surfaces. These gland cells may be demonstrated by their intense staining with Nile blue sulfate, neutral red, and Bismarck brown. They also stain with brilliant cresyl blue, crystal violet, thionin, and toluidine blue. These para-esophageal gland cells, whose function is unknown, are absent in cercariae of Echinostoma malayanum, E. hystricosum, Echiroparyphium dunni, and Hypoderaeum dingeri, but present in cercariae of an Echinostoma with 37 collar spines, obtained from a Brazilian Biomphalaria glabrata. Cercariae of E. audyi and E. lindoense, though closely related, are easily differentiated by the distribution of these para-esophageal gland cells and their outlets. Lie and Umathevy (1965) indicated that mature, free-living cercariae of Echinostoma audyi Lie and Umathevy, 1965 and E. lindoense Sandground and Bonne, 1940, have, in addition to cystogenous and penetration gland cells other unknown gland cells that open on the body surface and stain intensely in dilute Nile blue sulfate or neutral red. This paper presents further details about these unknown gland cells. Because of their location near the esophagus I propose to name them para-esophageal gland cells. MATERIALS AND METHODS Cercariae of E. lindoense were obtained from experimentally infected Gyraulus convexiusculus (Hutton) and those of E. audyi from Lymnaea rubiginosa (Michelin). The supply of these cercariae was unlimited as the parasites were maintained in the laboratory. Living cercariae were stained with Nile blue sulfate, neutral red, and other intravital dyes diluted in half-strength physiologic saline and examined under oil immersion. Lillie's (1954) periodic acid-Schiff sulfite leucofuchsin (PAS) stain was employed for cercariae fixed in 5% formalin, 70% and absolute alcohol. Received for publication 25 May 1966. * This work was supported by the University of California International Center for Medical Research and Training (Hooper Foundation, San Francisco School of Medicine) with Research Grant GM 11329 from the Office of International Research, NIH, U. S. Public Health Service and by grant No. MSC-Hooper Foundation-02-Trevey Fund from the University of California School of Medicine, Committee on Research, San Francisco Medical Center. Cercariae of Schistosoma mansoni Sambon, 1907 were used as controls.

Cell movement, proliferation and death in the formation of the embryonic axis of the rabbit.

AbstractRabbit blastocysts of ages six to eight days, post coitum were used to study the roles of cell proliferation, cell movement and cell death in the formation of the primary embryonic axis.Neutral red staining of local spots on the surface of cultured embryos provided information on the direction and speed of cell movement. Nile blue sulphate was used to detect necrotic cells in the embryonic areas. Mitotic indices were determined from counts made of the cells of serially sectioned embryos.It is concluded that the formation of the embryonic shield and primitive streak from the disc in the rabbit embryo is primarily achieved by migrations of cells that are being rapidly proliferated over the entire surface of the embryonic area. Cell death occurs but is an insignificant factor in this phase of embryonic growth.

Lipid of normal and regenerating limb tissues of the adult newt, Diemictylus viridescens: acidic and non-acidic lipids, phospholipids, and cholesterol.

AbstractCryostat‐cut sections of unamputated and amputated‐regenerating limbs of the adult newt were examined following the Nile blue test for acidic and non‐acidic lipids, the acid hematein and plasmal tests for phospholipids, and a Schultz test for cholesterol.Triglycerides (Nile blue test) are prominent in dermis and macrophages: triglyceride droplets are scattered in epidermis, wound epithelium, and regeneration blastema. Fatty acids (Nile blue test) are present in all tissues of the normal and regenerating limb: nerve myelin contains relatively little free fatty acid, while macrophages appear to contain the least amount of this lipidic substance.Plasmalogens (plasmal test) are prominent in the myelin of nerves, and macrophages: a weak cytoplasmic reaction obtains in the epidermis, subcutaneous glands, striated muscle, tunics of blood vessels, wound epithelium, blastema cells, chondrocytes, perichondrium and periosteum.Mitochondria responding for cephalin, lecithin, and sphingomyelin (acid hematein test) are ubiquitously distributed among the cells and tissues of the normal and regenerating limb. These phosphatides are prominent in nerve myelin, macrophages, and in dermal droplets: a variable response obtains from the myofibrils of striated muscle.Cholesterol (Schultz test) was demonstrated only in nerve myelin and in macrophages associated with injured nerves.Particular attention was paid to the lipid responses of the regeneration blastema, and the conclusion was reached that not all of the lipid previously demonstrated with sudan dyes was characterized by the current series of lipid tests.A modified Nile blue sulfate test that promises greater specificity in distinguishing between acidic and non‐acidic lipids is introduced.

The effects of triparanol on adrenocortical cells of the zona fasciculata of Syrian hamsters

Adrenocortical cells of the zona fasciculata of adult Syrian hamsters given 250 mg/kg triparanol daily by stomach tube were compared with cortical cells of untreated control animals. Dark cytoplasmic bodies which stained with toluidine blue, Nile blue sulfate, Sudan black B and acid hematein were observed with the light microscope. The bodies disappeared following the pyridine extraction-acid hematein test and control animals as well as animals treated with triparanol failed to give a positive reaction for cholesterol. These findings offer evidence that the cytoplasmic bodies are phospholipids. With the electron microscope the cytoplasmic granules exhibited rounded edges and were occasionally surrounded by a membrane. They displayed a crystalline latticework with opaque lines approximately 40–60 A separated by clear spaces 50–70 A. These observations coupled with the biochemical findings of other investigators suggest that the crystals may represent either the accumulations of enzymes or the matrix for a steroid precursor such as desmosterol.

The Life Cycle of Bunoderella metteri (Allocreadiidae: Bunoderinae), a Trematode Parasite of Ascaphus truei

The life cycle of Bunoderella metteri Schell, 1964, an intestinal parasite of the tailed frog, Ascaphus truei Stejneger in the Pacific Northwest, is described. Eggs are unembryonated when laid. Miracidia do not hatch in water. Ophthalmoxiphidiocercariae develop in rediae in the fingernail clam, Pisidium idahoense Roper. Larvae and pupae of the caddisfly, Rhyacophila grandis Banks, and dipterous larvae of the family Chironomidae serve as hosts for the encysted metacercariae. Immature specimens of B. metteri were recovered from the intestine of experimentally infected, laboratory-reared Rana aurora Baird and Girard, and Ascaphus truei. The only known bunoderid life cycle is that of Bunodera luciopercae (Miiller, 1776) Liihe, 1909, an intestinal parasite of freshwater fishes, reported by Wisniewski (1958). In Europe the cercariae develop in rediae in the freshwater clams Sphaerium corneum Linn. and S. rivicola Lamarck, and encyst in several species of Cladocera and Copepoda. Fishes become infected by ingesting the infected crustaceans. This paper presents the life cycle of an additional species. Bunoderella metteri Schell, 1964, is an intestinal parasite of the tailed frog, Ascaphus truei Stejneger, in the Pacific Northwest. Studies on its life cycle were undertaken concurrently in Idaho and Oregon. MATERIALS AND METHODS Adults of Ascaphus truei were collected at night from cold mountain streams in northern Idaho, western Oregon, and eastern Washington. The frogs remained alive many weeks when kept in water at 40 to 50 F. The water was changed every few days and saved as a source of parasite eggs that had passed in the feces of the host. Eggs collected by the sedimentation technique were placed in petri dishes on moist filter paper. Larval stages of the parasite were studied alive. Vital dyes (neutral red and Nile blue sulphate) were used to observe gland and flame cells of the cercariae. Naturally infected first intermediate hosts, Pisidium idahoense Roper, were collected from the same localities. The clams were kept alive for many weeks in dishes containing cold stream water and Received for publication 14 December 1964. * School of Natural Sciences, Arizona State College, Flagstaff. t Department of Biological Sciences, University of Idaho, Moscow. + Department of Zoology, Oregon State University, Corvallis. about 0.5 inch decaying vegetation. Measurements are in microns unless otherwise indicated.

THE CYTOLOGICAL DIAGNOSIS OF RUPTURED USING NILE BLUE SULPHATE STAINING.

BJOG: An International Journal of Obstetrics & GynaecologyVolume 72, Issue 3 p. 342-346 THE CYTOLOGICAL DIAGNOSIS OF RUPTURED MEMBRANES USING NILE BLUE SULPHATE STAINING I. Brosens M.D., I. Brosens M.D. Fellow of the Belgian Research Council (N.F.W.O.) Department of Obstetrics and Gynaecology, Hammersmith Hospital, London, W.I2Search for more papers by this authorH. Gordon M.B., F.R.C.S.(Ed.), M.R.C.O.G., H. Gordon M.B., F.R.C.S.(Ed.), M.R.C.O.G. Senior Registrar Department of Obstetrics and Gynaecology, Hammersmith Hospital, London, W.I2Search for more papers by this author I. Brosens M.D., I. Brosens M.D. Fellow of the Belgian Research Council (N.F.W.O.) Department of Obstetrics and Gynaecology, Hammersmith Hospital, London, W.I2Search for more papers by this authorH. Gordon M.B., F.R.C.S.(Ed.), M.R.C.O.G., H. Gordon M.B., F.R.C.S.(Ed.), M.R.C.O.G. Senior Registrar Department of Obstetrics and Gynaecology, Hammersmith Hospital, London, W.I2Search for more papers by this author First published: June 1965 https://doi.org/10.1111/j.1471-0528.1965.tb01468.xCitations: 31AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume72, Issue3June 1965Pages 342-346 RelatedInformation

SEMI-CONDUCTION AND ELECTRODE PROCESSES IN BIOLOGICAL MATERIAL. I. CRUSTACEA AND CERTAIN SOFT-BODIED FORMS.

The presence of semiconducting materials in a biological membrane across which a potential difference is established by ionic diffusion and active transport may be expected to lead to a leakage of charge through the electronically conducting elements. Oxidation and reduction will occur at the two surfaces which will therefore act as electrodes. In experiments with various Crustacea, particularly Palaemonetes varians (Leach) and Carcinus maenas (L.) the normal potential of the living animal is shown to be negative and, whether living or preserved, a negative potential is generated by diffusion when the salinity of the surrounding water is decreased. The presence of semi-conducting properties in the cuticle is shown by the occurrence of electrode processes following the application of potentials higher than those which exist in the normal animal. Thus when the surface is made negative hydrogen is liberated and metallic silver and copper are rapidly deposited from solutions of their salts. In the cuticle of living material at normal potentials or at potentials enhanced by outward diffusion of salt following transfer to reduced salinity an alkaline reaction is produced at the surface. This is shown particularly by the use of the dye nile blue sulphate, which is adsorbed by cathodic surfaces where it then acts as an indicator, alkalinity producing the red free base. Colours similar to those produced at the surface of negatively charged Crustacea appear at metallic cathodes. Alkalinity appears at the surface of living and non-living Crustacea in places where electrode processes are demonstrable at high potentials and comparable effects are not produced by diffusion in soft tissues or in gelatin and other substances in the absence of organic semi-conductors. The appearance of alkalinity at these positions is therefore attributable to cathodic reduction of salt water and not to the segregation of hydrogen and hydroxyl ions by diffusion processes. In distilled water the rate of reduction of ammoniacal silver nitrate on the cuticle is greatly increased by the action of a diffusion potential, showing that this reaction is here at least partly electrolytic in nature and that the surfaces on which deposition occurs are acting as cathodes. Similar phenomena can be seen in Cyanea and other planktonic coelenterates; thus with certain precautions electrolytic hydrogen may be evolved from the surface and in the living and undamaged animal nile blue adsorbed at the surface develops a mauve to purple tint similar to that of undamaged Crustacea, indicating a slightly alkaline reaction. In these forms also the surface is therefore acting partly as a cathode.

THE HISTOLOGICAL PROPERTIES OF RHODANILE BLUE.

(1) Rhodanile Blue is a compound of Rhodamine B (amphoteric) and Nile Blue Sulphate (basic). (2) Sections stained in a 1 per cent aqueous solution of it for half an hour show a polychrome picture in red, violet, blue. (3) Experiments invitro et in tela indicate that the colours produced by this dye reflect its activity as a redox substance, one that oxidises some tissue elements, reduces others, and is unaffected by others again. (4) The dye is a distinctive stain for fibrin as well as being a useful stain for the matrix of keratinous tissues, whose birefringence it intensifies somewhat.

Methemoglobin Reduction: Studies of the Interaction between Cell Populations and of the Role of Methylene Blue

Abstract1. It has been shown that mixtures of normal and G-6-PD deficient nitrite-treated erythrocytes reduced methemoglobin at a rate considerably more rapid than that computed from their individual rates of methemoglobin reduction.2. Differential agglutination studies demonstrated that when normal, methemoglobin-containing cells in such a mixture have reduced all of their methemoglobin, they facilitate methemoglobin reduction in G-6-PD deficient erythrocytes.3. The same effect could be observed in other mixtures of cells, (e.g., normal with normal, G-6-PD deficient with G-6-PD deficient, etc.) and even with highly purified methemoglobin solutions.4. This effect could be observed in the presence of methylene blue but not in the presence of another redox dye, Nile blue sulfate.5. Lactate served as an effective substrate for methemoglobin reduction. Methemoglobin reduction by lactate was enhanced by methylene blue but not by Nile blue sulfate.6. In mixtures of normal and G-6-PD deficient erythrocytes, no deficit in the rate of accumulation of lactate was found. This indicates that the mechanism of enhancement of methemoglobin reduction is not the diffusion of lactate from non-methemoglobin-containing cells to methemoglobin-containing cells.7. It was demonstrated that leukomethylene blue could reduce highly purified solutions of methemoglobin in the absence of the enzyme "methemoglobin reductase."8. The possible mechanism by which non-methemoglobin-containing cells may reduce methemoglobin in methemoglobin-containing cells is discussed. It seems most probable that leukomethylene blue is the mediator of the effect. This implies, contrary to earlier suggestions, that "methemoglobin reductase" acts prior to the reduction of methylene blue in the electron transport chain.

VITAL DYES FOR MARKING LIVING ELATERID LARVAE

The following dyes appear to be suitable as internal vital markers for living wireworms: Nile blue sulphate, rhodamine B, Sudan black B, brilliant cresyl blue, acridine orange N, coriphosphine O, and brilliant fat scarlet. All are assimilated quickly after ingestion with food, are stored in the fat body for extended periods, can be seen distinctly through the integument, and have no apparent toxic effects. Their selective storage in the fat bodies appears to be a product of their content of the fat-soluble amino-ethyl or amino-methyl groups, and of a specific type of molecular configuration. These dyes will undoubtedly also be useful as vital markers for other insects, particularly those with thin, transparent integuments.

On colouring epon-embedded tissue sections with Sudan black B or Nile blue A for light microscopy

ABSTRACT Sections of osmium-fixed tissues embedded in epon 812 colour with either Sudan black B or Nile blue A solutions to reveal a variety of detail by direct microscopy with normal apochromatic or semi-apochromatic objectives. The clarity of the coloration gives a picture fully comparable to that seen by phase-contrast microscopy. The plastic is not removed. 1μ sections or thinner sections down to green or gold, are mounted on clean glass slides by drying down from 20% acetone/water after flattening on a hot plate. Colouring is carried out at room temperature in Sudan black B (saturated solution in 70% alcohol) for 1 to 2 h. The result is a reversed or negative effect, for the epon plastic takes the stain avidly, but dense elements of the tissue do not, and appear white against a blue background of stained plastic. Lipid droplets retain a capacity to colour, becoming dark blue to blue-black. Nile blue sulphate (1% aqueous solution) colours thin sections of tissue in 1 to 2 h at 60° C, acting apparently as a basic dye on most cell elements, and also colours lipid droplets dark blue. After both techniques the sections are mounted in Farrants’s medium.

Chemocytological studies on a sudanophile substance in parathyroid cells of the toad (Bufovulgaris japonicus).

AbstractA sudanophile lipid was demonstrated in the parenchymal cells of the toad (Bufo vulgaris japonicus) parathyroid gland in specimens taken at the end of hibernation. A series of histochemical staining methods and extraction techniques for lipids were applied to the parathyroid tissues in an attempt to characterize chemocytologically the sudanophile substance. From its chemocytological properties such as sudanophilia and nile blue sulfate and acid hematein stainability, the substance was concluded to consist primarily of glycerides and phosphatides. The cytophysiological significance of these lipid moieties in the parathyroid cells was discussed in relation to the hibernation of the amphibian species.

CYTOLOGICAL STUDIES DURING GERMINAL VESICLE BREAKDOWN OF PECTINARIA GOULDII WITH VITAL DYES, CENTRIFUGATION AND FLUORESCENCE MICROSCOPY

1. Living eggs of Pectinaria gouldii were stained with vital dyes and vital fluorochromes before and after germinal vesicle breakdown. Observations were made with the bright field, dark field and fluorescence microscopes. Changes in the germinal vesicle, nucleolus and chromosomes of the living eggs were followed.2. Other eggs were vitally dyed and centrifuged in order to stratify the cell particulates. Cell granules were identified, based upon their specific staining and their stratified position in the centrifuged egg. These included lipid droplets, lipid granules, mitochondria, proteid yolk, basophilic and cortical granules. Limited cytochemical tests were made to verify their identity.3. Two kinds of metachromatic granules were seen. With toluidine blue, the granules occur at the centripetal pole just beneath the lipid cap. When Nile blue sulphate was applied, a different metachromatic band stratified just centripetal to the heavier yolk granules. Astral granules originate from the latter metachromatic band and became associated with the first maturation spindle.4. The fluorescent cell components included cell and nuclear membranes, nucleoli, chromosomes, lipid granules, two types of yolk granules and mitochondria.

Histochemical Studies in Primary Lipoidoses

Histochemical investigations were performed on the stored lipids in cases of infantile and juvenile amaurotic idiocy, gargoylism, Gaucher's disease and Niemann-Pick's disease.The results obtained are summarized as follows.1. A new histochemical detecting method of ganglioside was established.2. In amaurotic idiocy, cerebroside especially ganglioside and phosphatide especially sphingomyelin are deposited in the ganglion cells. In gargoylism, cerebroside especially ganglioside and phosphatide especially sphingomyelin are detected in the ganglion cells. Acid mucopolysaccharide is deposited in the perivascular tissue and probablly in the ganglion cells. In Gaucher's disease cerebroside is detected in the enlarged cells. In Niemann-Pick's disease, phosphatide especially sphingomyelin is deposited in the enlarged cells.3. The stored lipids in primary lipoidoses give positive reactions to all of sudan III, PAS and nile blue sulfate stains.4. For differentiations of stored lipids in primary lipoidoses, the test method for cerebroside, Bial's test, the common test method for phosphatide, alcian blue stain and Smith-Dietrich's method are believed to be useful.

Effects of Gamma, High-Energy Electron, and Thermal Neutron Radiations on the Fibrillar Structure of Cotton Fibers

Changes in the structure of cotton fibers after expesure to gamma, high-energy electron, and thermal neutron radiations, in various ambient atmospheres, were evaluated by the techniques of optical and electron microscopy. Radiation exposures evaluated were: gamma rays, 105 to 4 × 108 r.; 2-Mev electrons, 5 × 104 to 108 rep; integrated thermal neutron fluxes, 5 × 1014 to 1017 nvt. Changes in the physical and chemical properties of similarly irradiated fibers have been, or are to be, reported in other pub lications. All irradiated, unmodified cottons could be differentiated from unexposed cottons by optical microscopical observations of samples stained in Nile Blue Sulfate. Gamma ray and electron exposed samples differed from unexposed fibers. in swelling behavior in 0.2 M ( in Cu) cupriethylenediamine hydroxide, but thermal-neutron-irradiated cotton differed from the control only after exposure to an integrated flux of 1017 nvt. Electron micrographs of the fibrillation patterns exhibited by fibers exposed to gamma and electron radiation doses of 5 X 106 to 108 r. or rep on beating in water in a Waring Blender showed the formation of increasingly smaller fragments of samples. Some shortening of fibrils and an increase in number of broken ends of microfibrils were seen in neutron-irradiated cotton exposed to 10" nvt; at lower integrated neutron fluxes, fibrillation was unchanged. No distinct differences were found in microscopical tests between gamma and electron irradiated fibers or between gamma-exposed fibers irradiated in oxygen and nitrogen. At doses of 4 × 10" r. of gamma rays, the trend toward smaller fragments reversed and short chunks of material were observed. Varying amounts of degradation were observed in modified cottons after irradiation. Smaller fragments were observed in cottons modified to change and reduce the total crystalline cellulose; the fibrillation of acrylonitrile-treated cotton appeared unchanged, but irradiation produced a decrease in the size of fragments of dichloropropanol- and dimethylol cyclic ethyleneurea-modified samples.