Fluorescence of eosinophil leucocyte granules induced by 1-hydroxy-3,6,8-pyrenetrisulfonate. Visualization of differences in protein isoelectric points

After treatment of horse, rat and human blood smears with alkaline solutions of 1-hydroxy-3,6,8-pyrenetrisulfonate (HPTS), eosinophil leucocyte granules were the unique cell components which showed a bright green fluorescence. When stained with HPTS at pH 10, the whole granule of horse eosinophils showed high emission which strongly diminished after washing or staining in salt solutions or by using blocking methods for amino groups. Using HPTS at pH 12, the fluorescence reaction of house granules was specifically located in the peripheral region, appearing as fluorescent rings. These microscopic observations, which indicate differences in the isoelectric point of proteins within the eosinophil granule, were also confirmed by HPTS staining of protein blots as model substrates. Spectral analysis of HPTS at pH 10 and 12 showed practically identical absorption and emission spectra with peaks at 450 nm and 510 nm, respectively. Our results indicate that mainly ionic binding occurs between cationic proteins and HPTS in alkaline solution, and that the most cationic proteins (with isoelectric points at pH higher than 12) are located in the peripheral annular region of horse eosinophil granules.     

Viscosity decrease of pectin and fruit juices catalyzed by pectin lyase from Penicillium italicum in batch and continuous-flow membrane reactors

Summary The extracellular pectin lyase (PNL, E.C. 4.2.2.10) from Penicillium italicum was utilized in batch and confined in a continuous-flow ultrafiltration membrane reactor. The enzyme catalyzed the decrease in viscosity of pectin solutions at pH 6.0 as well as of different fruit juices at their respective pH. PNL decreased the viscosity of pectin solutions in the membrane (60% after 30 min) more than in the batch (46% after 70 min) reactors, but similar viscosity reduction of fruit juices was achieved in both reactors. The enzyme decreased the viscosity of grape, peach and melon juices to different extents, but failed to do so with apple or pear juices. It can be concluded that the utilization of PNL in a membrane reactor appears of interest for the clarification of fruit juices.     

Synthesis of new azino fused benzimidazolium salts. a new family of DNA intercalating agents. I

A series of new pyrido[1,2-a]- and pyridazino[1,6-a]benzimidazolium salts have been synthesized from readily available 1,3-disubstituted 2-alkylbenzimidazolium salts. Their affinity to DNA and in vitro cytotoxicity versus HT-29 have been tested. The initial results show that the title compounds are a new family of intercalating agents.     

Batch culture of Candida utilis in a medium deprived of a phosphorous source

Summary Candida utilis var. major NRRL-Y-1084 was grown in a defined medium without a phosphorous (P) source. During the exponential phase, cells divided according to a specific growth rate of 0.32 h^-1, which is lower than the usual rate for a balanced medium (0.4–0.6 h^-1). The relative P content of the biomass decreased from 2.70% to 0.75% over a period of 6 h, including 2 h of cell division arrest. At the end of this period there was another interruption of cell division. After that, multiplication restarted at a considerably lower rate and it deviated slightly from the exponential pattern. The stationary phase began when biomass P content reached 0.4%–0.5%, slowly decreasing afterwards to 0.25–0.20%. Biomass synthesis was less affected than cell division by the relative decrease of endogenous P, the two processes differing partially in their kinetics. Cell lysis started shortly before the stationary phase and affected about 20% of the population by the end of the assay. RNA and P content of the resulting biomass were 2.4% and 0.25% respecitvely, P being mainly incorporated to RNA.The relationship of biomass production to glucose uptake was very low, probably because the marked P deficiency called for an increase in energy consumption for growth and specially for maintenance. Compared with yeasts grown in a balanced medium, 40% increase in glycogen was observed, whereas no mean changes in the content of cell wall carbohydrates (glucan and mannan) and that of true protein were found.Member of the Scientific Researcher's Career of the Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET). Agrentina     

Microscopic anatomy of the ovary ofAlouatta caraya

The microscopic structure of theAlouatta caraya ovary is studied in different ages and reproductive stages. The most significant feature seems to be the presence in adult ovaries of abundant glandular interstitial tissue which occupies both the cortex and medulla. It seems to be derived from the theca interna of atretic follicles. Discrete luteinized masses are present in the medulla in all the ovaries observed. Invaginations of the surface epithelium are seen only in infant and juvenile ovaries. The development of cystic follicles seems to be a common pathway of atresia.     

Growth promotion and inhibition by antibiotic-producing fluorescent pseudomonads on citrus roots

Summary Forty-three strains of feeder root colonizing fluorescent pseudomonads from rough lemon (Citrus jambhiri Lush.) roots were examined for effects on rough lemon and sweet orange (Citrus sinensis Osbeck) seedlings. Plants inoculated with a single bacterial soil-drench had, after 10 months, a range of stimulatory (to 116%) and inhibitory effects (to 52%). Stimulatory bacteria particularly increased growth of root systems. Cultivar-specific inhibition and stimulation was evident in inoculations of rough lemon and sweet orange seedlings. Populations of fluorescent rhizobacteria on inoculated and noninoculated, as well as on stimulated and nonstimulated seedlings, did not differ significantly (10.8×10⁶ to 30.3×10⁶ CFU/g root). Population of fluorescent rhizobacteria on seedlings were higher than populations on feeder roots from grove trees (2.8 to 5.7×10⁶ CFU/g). Ninety-four and 81% of 251 fluorescent strains produced antibiotics against the fungusGeotrichum candidum and the bacteriumErwinia stewartii, respectively. Antibiotic activities of 90% of the antibiotic producing strains were repressed by Fe³⁺, indicating siderophore production. In comparison, only 9.6 and 15% of 94 randomly selected nonfluorescentPseudomonas strains were antibiotic producers. Differences between stimulatory and inhibitory or neutral bacteria were not apparent from antibiosis tests. On the basis of physiological tests,Pseudomonas putida was the most abundant (>62%) pseudomonad species on rough lemon roots. Growth stimulating strains appeared to be in bothP. putida andP. fluorescens groups. FewP. aeruginosa strains were identified on citrus roots.Florida Agricultural Experiment Stations Journal Series No.     

Qualitative dynamics of three species predator-prey systems

Summary A qualitative analysis of some two and three species predator-prey models is achieved by application of the method of averaging in conjunction with a Lyapunov function constructed from the appropriate Volterra-Lotka model. We calculate the limit cycle solution for a two-species model with a Holling type functional response of the predator to its prey by means of a time-scaled transformation. The existence of a bifurcation of steady states for a community of three species is discussed and the periodic solution around one of the unstable steady states is calculated to the lowest approximation. Several comments are made regarding the behavior of these systems under changes of some control parameters.This work was supported in parts by USERDA, Contract number E(11-1)-3001.     

An Artifact Of Liquid Emulsion Autoradiography

The localization of ³H-opiatcs in the myenteric plexus of the guinea pig ileum is subject to systematic artifact when stretch preparations of the myenteric plexus are dipped into liquid Kodak NTB-3 emulsion for autoradiography. The cause of the artifact was determined to be a discontinuous distribution, or retraction, of emulsion over plexuses. The apposition of frozen freeze-dried ilial sections to dried photographic emulsion avoids this source of error.