Role of cell-mediated immunity in the resistance to experimental sporotrichosis in mice

The in vitro activity of several new imidazoles, cloconazole, sulconazole, butoconazole, isoconazole and fenticonazole, were compared with those of amphothericin B, flucytosine, and three azoles: econazole, miconazole and ketoconazole against isolates of pathogenic Candida. A total of 186 clinical isolates of 10 species of the genus Candida and two culture collection strains were tested by an agar-dilution technique. Isoconazole was the most active azole, followed by butoconazole and sulconazole. Differences between some of the species in their susceptibility to the antifungal agents were noted. Sulconazole and cloconazole had the highest activity in vitro against 106 isolates of C. albicans. Butoconazole and isoconazole were also very active against isolates of C. albicans, and were the most active azole compounds against 80 isolates of Candida spp.     

Rapid osmotic adjustment by a succulent halophyte to saline shock

The objective of this research was to measure the short term osmotic adjustment of Salicornia europaea L. ssp. rubra (A. Nels) Breitung when suddenly exposed to 100 millimolar NaCl. Plants were grown hydroponically, shocked with 100 millimolar NaCl added to the culture solution, and stem tips analyzed for free inorganic ions and small organic molecules at intervals up to 72 hours. In the first 2 hours, the calculated leaf osmoticum showed a net increase of 158.8 millimolar most of which was free Mg²⁺ (+135.3 millimolar). Total sugars increased almost 5-fold by the 6th hour, enough to provide sufficient osmoticum for the cytoplasm if only partially confined there. By 24 hours, all measured osmotica had decreased except Na⁺, Mg²⁺, Cl⁻, and proline, with the net increase being 208 millimolar. By 72 hours, there was a net gain of 356 millimolar in osmotica of the stem tips, due to Na⁺ (+233.3 millimolar), Cl⁻ (+306.7 millimolar), and a small increase in sugar and proline (+3.5 millimolar), with all other osmotica decreasing in concentration. Compatible osmotica did not change sufficiently to account for osmotic balance between vacuole and cytoplasm; consequently, there must have been a reapportionment of osmotica within the cell in the short time duration of this experiment.     

DETERMINATION OF GUSTATORY SUCROSE THRESHOLD IN WATER BY USE OF A LINEAR SUCROSE GRADIENT

A novel experimental method was developed which allows the determination of the threshold concentration of sucrose by use of a linear sucrose gradient in water. With this method a continuous tasting of the test-liquid is possible. A panel of 15 persons experienced in taste-testing was used. Three gradients of different steepness were applied: 0 to 1.5% (w/w) sucrose in 2 min (I), 3 min (II) and 4 min (III). The results of the new method were compared with those of the standard method (DIN). With gradients I and II we found values which were significantly higher than those of the standard method (I: 0.49% (w/w); II: 0.46% (w/w); DIN: 0.31% (w/w)), whereas with gradient III the same threshold value was found as with the DIN-Method (III: 0.32% (w/w)).     

The quantitative analysis of endogenous folate catabolites in human urine.

In man folates are catabolized and excreted as inactive cleaved degradation products, a mixture of pteridines and p-aminobenzoylglutamate (pABGlu) or its acetamido derivative (apABGlu). The daily rate of excretion represents the inescapable use of the vitamin in metabolic activity and thus has implications for determining the recommended dietary allowance for the vitamin. Furthermore, the rate of catabolism has been suggested to rise during pregnancy and in certain disease states. A method is described for the quantitative extraction and assay of the folate catabolites pABGlu and apABGlu in human urine. Aliquots of 24-h urine collections are acidified and applied to columns of Dowex 50W cation-exchange resin. The catabolites are selectively batch-eluted with increasing concentrations of HCl. The fraction containing pABGlu is diazotized and then applied to a C18 Sep Pak column for further purification and concentration. The fraction containing apABGlu was deacetylated and reapplied to the Dowex column and then treated identically to the pABGlu fraction. The methanolic concentrates of both extracts were evaporated to dryness and reconstituted with water and pABGlu was regenerated by reductive cleavage of the diazotized material with Zn/HCl. The extracts of the two catabolites were separated by reverse-phase HPLC using a Radial Pak C18 column. Recovery of isolated material was monitored by the addition of high specific activity tritiated labels of both compounds added as internal standards to all urine aliquots prior to purification and analysis.     

Fine structure of the pineal organ in the troglobytic fish,Typhlichthyes subterraneous (Pisces: Amblyopsidae)

Summary The pineal organ of the blind, cave-dwelling fish, Typhlichthyes subterraneous, was examined with both light and electron microscopes. Like the eyes, the pineal in this troglobytic species was found to be regressed. Two cell types, photoreceptor and supportive cells, were described in the pineal epithelium. Although ganglion cells were not identified, small, unmyelinated nerve fibers were present. The photoreceptor cells had degenerated outer segments. Accordingly, it was suggested that the pineal in this species is not likely to function in photoreception. However, the presence of well developed Golgi bodies, clear and dense-cored vesicles, variable amounts of rough endoplasmic reticulum and glycogen particles indicated that both cell types are metabolically active and may play a role in secretion.     

应用气-质联用仪测定三种螺旋藻中的甾醇成分

应用GLC/MS联用仪对室内培养的钝顶螺旋藻(Spirulina platensis (Nordstedt) Geitler)、极大螺旋藻(S.maxima (Stechell & Gardiner) Geitler)和盐泽螺旋藻(S.subsalsa Oerst)的甾醇成分进行了测定。从钝顶螺旋藻和盐泽螺旋藻中共分出11个相同的甾醇组分:胆甾醇、胆甾烷醇、芸苔甾醇、麦角甾醇、海绵甾醇、菜子甾醇、豆甾醇、24-乙基-Δ~(5,7,22)-胆甾醇、β-谷甾醇、异岩藻甾醇和4α,23,24-三甲基Δ~(5,22)-胆甾醇;从极大螺旋藻中只分离出8个甾醇组分。其中胆甾醇含量最高。4α,23,24-三甲基-Δ~(5,22)-胆甾醇为蓝藻中首次报导。     

Ultrastructural observations on synaptic ribbons in the pineal organ of the goldfish

Summary Synaptic ribbons in the pineal organ of the goldfish were examined electron microscopically with particular attention to their topography. These structures were formed of parallel membranes, which were poorly preserved with OsO₄ fixation and could be extracted from thin sections with pronase indicating their proteinaceous nature. Synaptic ribbons were closely apposed to the plasma membrane bordering dendrites of ganglion cells, but were also related to processes of both photoreceptor and supportive cells. Their close proximity to invaginations of the plasma membrane and portions of the endoplasmic reticulum suggest that they are involved in the turnover of cytoplasmic membranes. Tubular and spherical organelles of unknown function are also described.     

Dual labeling in standard DNA-RNA hybridization studies using 125 I-labeled nuclear RNA and 3H-labeled DNA.

Standard DNA-RNA hybridization studies, using nucleic acids isolated from mammalian tissues, are frequently hindered by relatively low levels of radioactivity in pulse-labeled RNA and in an inability to reliably estimate the amount of DNA present in the hybrid. In the method described here nuclear RNA is labeled in vitro with 125I to 400 000- 800 000 cpm/mug and DNA is obtained from a rat glial tumor line grown in culture and labeled to specific activities of 42 000-79 000 cpm/mug. DNA-RNA hybridization is conducted in an all solution system at RNA:DNA ratios of 3.5:1 to 18:1. Assay background is controlled by pretreatment of the hybrid and free RNA at the conclusion of the annealing study with RNase, then isolation of the hybrid together with a small fraction of free RNA oligonucleotides on hydroxyapatite. The partially purified hybrids are then trapped on Millipore filters. Assay background id 0.004% of total counts present in the annealing reaction. Comparison of the annealing reactions of pulse-labeled liver nuclear RNA and in vitro 125I-labeled nuclear RNA in saturation, kinetic, and competitive hybridization studies shows them to be essentially the same. Nuclear RNA labeled by either tritium or iodine shows a 10-20-fold greater concentration of the annealing sequences over that found in the microsomal RNA. Minor differences are noted between the nuclear RNAs in the initial rates of reaction and in the magnitude of the decrease in percent hybridization at low levels of unlabeled competitor RNA. This may be due to preferential labeling in pulse-labeled RNA of molecules which are present in lower concentrations or are transcribed from more frequently repeated DNA sequences than the average population of annealing RNA molecules. The technique has application in systems where the amount of tissue for RNA extraction is small or where the system does not permit the obtaining of pulse-labeled RNA, as in experimental rodent skin carcinogenesis or in dealing with RNA from the tissues of large mammals or humans.