Berberine Sulfate Research Articles

Pharmacological Investigations of Berberine Sulphate

The alkaloid, berberine, isolated from Berberis aristata Linn has been used in the Indian system of medicine as a stomachic, bitter tonic and also in the treatment of oriental sores. The root bark and its decoctions were thought be an excellent remedy against malaria and were used as a diaphoretic and antipyretic in the past (1). It was reported to possess bacteriostatic activity especially against V. Cholera and was found to be useful in the treatment of cholera and diarrhoea (2). Recently, it has been claimed that berberine and its salts have a place in the chemotherapy of amoebiasis (3-4). The present work is devoted to the study of the antiamoebic properties of this drug on experimentally induced amoebiasis in rats, the minimum inhibitory concentration and other relevent pharmacological effects on experimental animals and animal tissues.

Fluorescent staining of juxtaglomerular cells in frozen sections.

Enzymatic investigations of the juxtaglomerular apparatus often creates the need for visualisation of granulated juxtaglomerular cells (JGC) in preparations subjected to histochemical procedures. In our investigations, Pitcock and Hartroft's (1958) modification of Bowie's method and the Endes et al. (1969) combined trichrome staining proved to be inadequate when applied to fresh cryostat sections, or to formol- or glutaraldehyde-fixetl, gum sucrose-impregnated frozen sections. Friedberg and Reid's (1966) crystal violet procedure for waxembedded kidneys also failed to give uniformly reproducible results. In attempting to find a satisfactory technique for both enzyme and granule staining, we noted Janigan's (1965) and Haratla's (1969) observations on paraffin-embedded JGC, and tested the following fluorochromes: thioflavine T—Fluka, C. I. 49005; auramine O—Merck, C. I. 41000; acridine orange—E. Gurr, C. I. 46005; berberine sulfate—Fluka, C. I. 75160 on 10 μ sections of albino mouse kidneys prepared in 4 dif...

Some methods for fluorochromation and staining of rat mast cells with bacic dyes.

Abstract— Fluorochromation and staining possibilities of rat mast cells with basic dyes have been studied. The material consisted mainly of tongue, salivary and lymph glands, as well as decalcified material from jaws containing incisors. A very selective fluorescence for mast cells was achieved with several basic fluomchromes, but berberine sulfate, acridine orange, and thioflavine T proved especially useful. Some combinations of dyes to be used for ordinary and/or ultraviolet light are described. The most useful seemed to be acridine orangelhematoxylinleosin, acridine orangelaniline blue WS, berberine sulfatelthiazine red R. The acid dyes mainly served as background stains. Good results were achieved in ordinary light with the basic dye methyl green, which also revealed interesting color change phenomena (“allochromasia”).

A Test for Stink Bug and Fungal Damage in Green Rice12

Tests were made using hydrochloric, acetic, and carbolic acids, hydrogen peroxide, Lugol’s solution, acid fuchsin, and berberine sulfate stains, and X-ray radiograph for treating shelled green rice to determine the extent of Slink bug, Oebalus poecilus (Dallas), and fungaldamaged kernels, Procedures included autoclaving, boiling, and heating the rice in various concentrations of the chemicals. Hydrochloric, acetic, and carbolic acids in various concentrations and trcalmenls turned damaged areas dark but did not affect undamaged areas of the kernels. Best results were achieved when the green rice was placed in hydrochloric acid in dilutions 0.03-15 N heated to 54°C for 1 minute. This treatment resulted in good contrast between damage and undamaged areas and was quick, easy, cheap, reliable, and free from the unpleasant odors of acetic and carbolic acids.

Berberine sulfate: antimicrobial activity, bioassay, and mode of action

Berberine sulfate was shown to possess antimicrobial activity against a wide variety of microorganisms including Gram-positive and Gram-negative bacteria, fungi, and protozoa. The antibacterial activity against Vibrio cholerae and Staphylococcus aureus was dependent on the inoculum size of the test organism and pH of the medium. A method of microbiological assay sensitive to 5–10 μg/ml of the drug was developed. The drug was shown to exert a more rapid antibacterial activity than chloramphenicol and tetracycline on V. cholerae, the K values being 2.4 ×10−2 sec−1, 7.8 × 10−3 sec−1, and 5.2 × 10−3 sec−1 respectively. Berberine sulfate was shown to be bacteriocidal to V. cholerae and bacteriostatic to S. aureus, at concentrations of 35 and 50 μg/ml. In both these organisms concentrations of 35 and 50 μg/ml of the drug inhibited ribonucleic acid (RNA) and protein synthesis almost immediately after the addition of the drug. There was little effect on deoxyribonucleic acid (DNA) synthesis at these concentrations.

Staining juxtaglomerular granules with basic fluorescent stains.

Many basic fluorescent dyes stain juxtaglomerular granules to produce characteristic colors in ultraviolet light. The stain is applied to paraffin sections of tissues fixed in 2% calcium acetate-10% formalin or in phosphate-buffered 10% formalin. Procedure: Bring section to water, stain 0.5 min in Delafield hematoxylin, wash in tap water, stain 3 min in a 0.1% aqueous solution of basic fluorescent dye (auramine O, acriflavine, acridine orange, coriphosphine O, acridine yellow, phosphine E, thioflavine T, berberine sulfate, atebrine or rivanol) and differentiate 1 min in 0.1% acetate acid (or omit this step). After washing in tap water, air dry with or without subsequent mounting in a resin. Juxtaglomerular granules stain bright fluorescent yellow or orange against a dark background.

Ultrastructural demonstration of cartilage acid glycosaminoglycans

A method for the demonstration of cartilage acid glycosaminoglycans by light and electron microscopy is described. Rabbit ear cartilage was fixed in cacodylate buffered 2.5% methanol-free formaldehyde with 0.001 M Ruthenium Red andp-chloromercuribenzoate (PCMB). Dehydration was carried out in ethylene glycol followed by embedding in the water-soluble glycol methacrylate (GMA). In some experiments unfixed cartilage was rapidly dehydrated. Sections, 1 μ thick, and ultrathin sections from the same blocks were stained with 0.001 M Ruthenium Red. Semi-thin sections from cartilage fixed without heavy metal additives were, in addition, stained with the acidophilic fluorochrome Berberine sulphate. It was found that Ruthenium Red intensely stained the same pericellular zone that stained metachromatically with Toluidine Blue or fluoresced after staining with Berberine sulphate. Prior treatment with 0.05% cetylpyridinium chloride entirely blocked the three reactions. Previous digestion with 0.2 mg hyaluronidase/ml for 30 min at 37°C led to the abolition of the fluorescence reaction with Berberine sulphate. It is concluded that Ruthenium Red selectively stains cartilage acid glycosaminoglycans. With the electron microscope the pericellular zones were found to be built up of a three-dimensional branched meshwork of fibrils covered with a mantle of electron-dense material, presumably acid glycosaminoglycans bound to Ruthenium Red.

Estimation of Messenger RNA in Fern Gametophytes

Spores of Equisetum arvense and Osmunda japonica, and protonemata of Pteris vittata were cultured with actinomycin D and with chloramphenicol. As a result, the following was revealed.1) Equisetum and Osmunda spores can germinate normally in presence of these drugs at concentrations lower than 100μg/ml. Two-dimensional growth is arrested at or higher than 50μg/ml of actinomycin D and 10μg/ml of chloramphenicol, while one-dimensional growth is inhibited at higher concentrations.2) In Pteris protonema, as well as in Equisetum and Osmunda, actual arrest of the growth by actinomycin D occurs after the plant continued development of a certain stage for several days. This implies that messenger RNA is taking part in protein synthesis for several days. This is supported by the fact that the cease of growth comes earlier if the plant is cultured with chloramphenicol.(3) Localization at high concentration of RNA seems to be a key factor for two-dimensional growth. Such RNA is high-polymer other than soluble RNA, as verified by acquisition of fluoresence combined with berberine sulfate. It is retained for several days even if the growth was arrested by actinomycin D. Therefore, most of such RNA may consist of ribosome RNA. As such RNA cannot take part in two-dimensional growth by itself when applied with actinomycin D, the role of messenger RNA seems to be indispensable for the growth.

A Fluorescence Microscopic Study of the Cytoplasmic Granules of <i>Hydrodictyon reticulatum</i>

The spherical cytoplasmic granules of Hydrodictyon reticulatum of 0.5-1.0μ in diameter were stained by Nile blue and showed an intense fluorescence when excited by long wave length ultraviolet light. When these granules were stained with berberine sulfate and excited, they did not show any fluorescence. Therefore, these granules are different from the so-called spherosomes of Perrier since they showed no fluorescence after staining with berberine sulfate. The ground substance of the cytoplasm showed an intense yellow fluorescence after staining with berberine sulfate. Differential centrifugation showed that this fluorescence of the cytoplasm is emitted from the microsomes. In this study, spherical cytoplasmic granules as well as microsomes were distinguished from other cell inclusions by fluorescence analysis with these two dyes.

Interaction between nucleic acids and berberine sulfate.

Interaction between nucleic acids and berberine sulfate.

A Fluorescent Dye Method for Foliar Penetration Studies

lish the location of the penetrant on the cellular level. Histochemical techniques have been employed with great success by Knight et al. (10), Rohrbaugh (14), Rice and Rohrbaugh (13), and others to demonstrate the sites of entry and routes of movement of oils. In the case of aqueous solutions, on the other hand, most evidence is indirect in nature, and histochemical evidence is limited to very few reports such as those of Palmiter et al. (12) and Wylie (24) for penetration and movement of iron. Many workers (3, 5, 15, 21, 22, 23) have compared stomatal with cuticular penetration by such indirect methods as measurement of plant response to growth regulators or nutrients, autoradiography or counting of tissues containing radioactive tracers, or measurement of nonabsorbed chemical. The importance of stomatal entry has not been clearly established by these methods. More direct evidence is obtained by using other techniques. For example, Aslander (2) and Crafts (6) studied the penetration of dilute sulfuric acid solutions by microscopic detection of injury resulting from the toxic action of the penetrant; however, conflicting observations were reported. Autoradiography of leaf sections has been used by Gustafson and Schlessinger (9) to demonstrate penetration of cobalt60 into bean leaves in the dark, but to date this promising method has not been widely applied in attempts to determine the route of penetration. Because most of the conventional methods yield only indirect evidence, a more direct method involving a fluorescent dye has been testedl. This type of tracer was used to study foliar absorption of dew water by Steubing (18), who followed entry and movement of berberine sulfate with the fluorescence microscope. By a similar method, Butterfass (4) observed foliar penetration via the club-shaped hairs of Vicia faba L. and Phaseolus coccineus L. Orgell3 used both fluorescent and nonfluorescent dyes to study penetration of isolated plant cuticles. In the work reported here, the fluorescent tracer method has been considerably expanded and improved for application to the problem of foliar absorption of aqueous solutions.

Use of ultraviolet light and fluorescent dyes in bacterial counting.

Summary1. The use of ultraviolet light and fluorescent dyes as an adjunct to the Petroff-Hausser bacterial counting chamber technique is reported.2. This method has made bacterial counts more accurate because of clarity, ease, and rapidity.3. Berberine Sulfate was the dye of preference although Auramine-O and Rhodamine-B were gauged equally as good.4. Twelve strains of bacteria, one protozoa, and one fungi were used in testing this procedure.5. All 14 organisms gave excellent fluorescent reaction to 3 dyes and 11 strains gave excellent results to a fourth dye.

Demonstration of Globi and Leprosy Bacilli by Fluorescence Microscopy.

Hagemann1 reported the superiority of a fluorescence method using berberine sulfate stain for demonstrating leprosy bacilli in nasal mucus and thick blood smears. The same year he2 proposed an auramin-fluorescence technic for the routine identification of acid-fast fast bacteria, especially tubercle bacilli. The suitability of this method for cultures of Myco. leprae has been confirmed by Kuster3 and for leproma smears by Kline and Leach.4Recently one of us (H.J.H.) has investigated the separation and concentration of acid-fast organisms from the tissues of leprosy patients. To cryochemed spleen† distilled water was added and direct impression smears were made from a piece of the tissue. One set of smears was stained with auramin O and examined according to the fluorescence procedure described by Richards and Miller.5A representative field is shown in the accompanying photomicrographs. The faint material in the background is non-acid-fast splenic cellular substance. The acid-fast bacilli have a characteri...

Equine Cutaneous Leishmaniasis: Treatment with Berberine Sulphate

Equine Cutaneous Leishmaniasis: Treatment with Berberine Sulphate