Localization of Endo-Oligopeptidase (EC 3.4.22.19) in the Rat Nervous Tissue

The subcellular and regional distribution of endo-oligopeptidase (EC 3.4.22.19), an enzyme capable of generating enkephalin by single cleavage from enkephalin-containing peptides, was determined by an enzymatic assay using metorphamide and by immunochemical techniques in the CNS of the rat. The rat CNS contains a membrane-associated form of endo-oligopeptidase, an enzyme predominantly associated with the soluble fraction of brain homogenates. Subcellular fractionation showed that approximately 17% of the total activity of the enzyme is associated with membrane fractions including synaptosomes. Synaptosomal membranes were prepared from neocortex, striatum, hypothalamus, medulla, spinal cord, and cerebellum. The amount of EC 3.4.22.19 activity solubilized by 3-[( 3-cholamidopropyl]dimethylammonio)-1-propanesulfonate from synaptosomal membranes was similar in neocortex, striatum, and hypothalamus, being three- to 10-fold greater than in spinal cord, cerebellum, and medulla. A polyclonal antibody exhibiting high affinity for endo-oligopeptidase was raised in rabbits against the purified rat brain enzyme and used to localize endo-oligopeptidase by Western blotting and by immunoperoxidase techniques. A strong band corresponding to the Mr of EC 3.4.22.19 was found in solubilized proteins obtained from synaptosomal membranes prepared from hypothalamus, neocortex, and striatum when subjected to Western blotting. The immunohistochemical localization of endo-oligopeptidase indicated that the immunoreactivity was confined to gray matter in regions known to be rich in peptide-containing neurons such as the striatum. In the cerebellum, a region poor in peptides, no staining could be detected. The nonuniform distribution of endo-oligopeptidase in rat brain suggests a role in neurotransmitter processing in the CNS.     

Multiple Bacteriocin Production by Leuconostoc mesenteroidesTA33a and Other Leuconostoc/Weissella Strains

Leuconostoc (Lc.) mesenteroides TA33a produced three bacteriocins with different inhibitory activity spectra. Bacteriocins were purified by adsorption/desorption from producer cells and reverse phase high-performance liquid chromatography. Leucocin C-TA33a, a novel bacteriocin with a predicted molecular mass of 4598 Da, inhibited Listeria and other lactic acid bacteria (LAB). Leucocin B-TA33a has a predicted molecular mass of 3466 Da, with activity against Leuconostoc/Weissella (W.) strains, and appears similar to mesenterocin 52B and dextranicin 24, while leucocin A-TA33a, which also inhibited Listeria and other LAB strains, is identical to leucocin A-UAL 187. A survey of other known bacteriocin-producing Leuconostoc/Weissella strains for the presence of the three different bacteriocins revealed that production of leucocin A-, B- and C-type bacteriocins was widespread. Lc. carnosum LA54a, W. paramesenteroides LA7a, and Lc. gelidum UAL 187-22 produced all three bacteriocins, whereas W. paramesenteroides OX and Lc. carnosum TA11a produced only leucocin A- and B-type bacteriocins. Received: 11 April 1997 / Accepted: 10 June 1997     

Guaraná (Paullinia cupana) as a commercial crop in Brazilian Amazonia

Guaraná (Paullinia cupana) is a woody vine or sprawling shrub native to the central Amazon Basin. The seeds are commercially produced on some 6,000 ha in the state of Amazonas near Manaus. The principal article of commerce is an amber-colored, carbonated soft drink. It is also widely used as a high caffeine stimulant and in local medicines. The plant is monoecious and is damaged by a number of diseases, the most severe being anthracnose. Prospects are excellent for greatly expanded international markets.     

Suppression of the rotifer Polyarthra remata by the omnivorous copepod Tropocyclops extensus: predation or competition

Indirect evidence from observations in the field suggested thata common and often abundant cyclopoid copepod, Tropocyclopsextensus, despite its small size (0.5 mm) and largely algaldiet, is an important predator of the rotifer Polyarthra remata.Laboratory experiments showed that copepodids and adults, butnot nauplii, markedly suppressed the population growth of thisrotifer. The suppression could be attributed entirely to predation,rather than to exploitative competition for shared food resourcesor to interference. Mortality rates predicted from a separatefeeding-rate experiment weresufficient to account for the declinein population size observed in the Tropocyclops treatment ofculture experiments. Also, monitoring the availability of cryptomonadfood in treatments with and without Tropocyclops throughoutthese experiments showed that cryptomonad densities always remainedhigh and were just as high or significantly higher in the copepodtreatments. Furthermore, direct videographic observations ofP. remata from cultures with and without Tropocyclops demonstratedno significant differences in swimming velocity, tendency todeviate from a straight-line path, or frequency and length ofspontaneous escape responses. Per capita ingestion rates ofT. extensus on P. remata varied from 2 to 8 day^-1, were significantlyhigher for adult females than adult males or copepodids V, andincreased significantly with rotifer density. A tendency ofcopepods to have higher ingestion rates on young than adultrotifers was not significant. Copepods cultured since birthwith cryptomonads and P. remata ate significantly fewer rotifersthan those cultured only with cryptomonads; this may be explainedby a more sated condition of the former and provided no evidencefor the idea that previous experience with this rotifer mightincrease predation efficiency. The results show that T. extensusin natural communities has the potential to deplete naturalpopulations of susceptible rotifer prey. Accordingly, it mayshift the species structure of rotifer assemblages in favourof resistant species, and provide selection pressure for avoidanceresponses.     

Hydrolysis of sunflower oil by means of hydrophobic membrane with lipolytic activity

Polytetrafluoroethylene (PTFE) membranes of different porosity were used for lipase from Rhizopus immobilization. The immobilized enzyme applied to sunflower oil hydrolysis achieved the activity of 1228 U/m² of the membrane area and the half-life time was calculated to be 7 days.     

Effects of oxygen and chloramphenicol on Frankia nitrogenase activity

The kinetics of asymbiotic nitrogenase activity in three strains of the actinomycete Frankia were studied. Decay rates for enzyme activity were determined by adding chloramphenicol to active acetylene-reducing cells and measuring the time required for all activity to cease. Synthesis rates were measured by bubbling oxygen through actively-reducing cells (which totally destroyed all activity) and then measuring the time required for activity to return to normal. Decay rates (t _1/2) for these three strains were approximately 30 to 40 min. Synthesis rates were slower and initial nitrogenase activities were recorded about 110 min (DDB 011610) or 210 min (DDB 020210 and WgCc1.17) after return to air-equilibrated cultures. Frankia strain WgCc1.17 showed a greater sensitivity to oxygen and nitrogenase activity was totally lost when cells were bubbled only with atmospheric concentrations of oxygen. The results presented here indicate that nitrogenase activity turnover time is relatively rapid, on the order of minutes rather than hours or days. However, regulation of nitrogenase activity will differ from one strain to another and asmmbiotic characterization will be useful for understanding nitrogenase regulation in the bacterial-plant symbiosis.Contribution no. 879 from the Battelle-Kettering Laboratory     

Different structure of polytene chromosomes ofPhaseolus coccineus suspensors during early embryogenesis

Summary The pattern of DNA and RNA puffs in pair VII of polytene chromosomes has been investigated in the suspensor ofPhaseolus coccineus during early embryo development. The pattern of³H-TdR and³H-U incorporation has been also detected. Collected data indicate that: 1. both heterochromatic regions, p₁₁ and q_(111+112), of chromosome pair VII, organize large DNA puffs; 2. DNA puffs of both regions are specific of different embryo differentiation steps; 3. a seasonal influence on the DNA puffing seems also to be present, as demonstrated by the comparison of the results collected in two different crops; 4. the incorporation experiment by³H-TdR evidences that not all DNA puffs show clustered labeling; 5. the RNA puffing of the two regions seems also to be specific of determined embryo stages.     

Reduction of a Tetrazolium Salt in Determining Growth Activity of Yeast-Phase Histoplasma capsulatum

A colorimetric method using a tetrazolium compound, 3,4,5-dimethylthiozalil-(1 or 2),2,5-diphenyltetrazolium bromide (TTBr), was developed for studying the growth activity of yeast-phase Histoplasma capsulatum. Materials extracted in phosphate buffer, pH 7.0, from cells at different stages of growth reduced TTBr. Colorimetric changes were correlated with enzymatic activity. Under standardized conditions specified herein, the optical density of the reduced tetrazole was an index of the growth activity of the organism.     

Studies on several genetic hematological traits of the Mexican population. XII. Distribution of blood group antigens in twelve Indian tribes

Blood samples from 1090 Mexican Indians belonging to the Chol, Chontal, Totonac, Huastec, Mixe, Mazatec, Zapotec, Mixtec, Chinantec, Nahua, Cora and Huichol linguistic groups, were obtained and examined in regard to the following blood group antigens: A, B. M, N, P, C, c, D, E, e, Fy(a), K and Di(a). The gene frequencies were similar to what has been described for other Amerindians; high values for O, M, CDe, cDE and Duffy; low to absent Kell and presence of Diego in variable amounts. The frequency of chromosomes CDE and cDe/cde was somewhat higher than usual and some of the tribes had relatively high frequencies of the A and B antigens. It was felt that variable degrees of non-Indian admixture was at least partially responsible for this situation. A previous study dealing with the distribution of abnormal hemoglobins and glucose-6-phosphate dehydrogenase deficiency in these same tribes, had strongly suggested the possibility of some Negro admixture in the Chontal, Nahua and Cora tribes. However, this was not specifically reflected in their blood group distribution. This served to emphasize the need of investigating as many markers as possible when trying to characterize a population.     

Effect of Drying on Unexposed Autoradiographic Emulsion in Relation to Background

Unexposed blanks prepared from Kodak AR 10 stripping film were dried by 3 methods: (1) fast, open drying with a fan for 25 min, (2) slow drying in a desiccator for 6 hr, and (3) very slow drying in a desiccator for 24 hr. The number of background grains depended on the mode of drying. Fast drying (method 1) gave 0.7 grain per 100 μ², slow drying (method 2) gave 0.33 grain; very slow drying (method 3), only 0.17 grain. The increase of background after fast drying is assumed to be caused by the rapid shrinkage of wet emulsion. This causes an increase in the intraemulsion pressure which, in turn, sensitizes the silver bromide crystals to cause an increase in the number of developable grains.