Application of pattern recognition to monitoring fermentations ofBacillus amyloliquefaciens

Summary Pattern recognition techniques were applied to analytical data to distinguish abnormal from normal microbial fermentations usingBacillus amyloliquefaciens as a model system. Patterns of fermentation end products during growth ofB. amyloliquefaciens were obtained from HPLC analysis of broth samples. Data were also obtained from fermentations using other bacterial species, strains, and environmental conditions, and were compared with the model data set. The bacterial species cultured includedB. subtilus, B. licheniformis, andEscherichia coli. Environmental variables included acration and temperature. The chromatographic patterns were compared by using hierarchical cluster and principal component analysis to obtain a quantitative measure of their similarity and to establish the normal variability within a model data set. Statistical analysis of the data indicated that individual fermentations can be assigned to distinct clusters on the basis of their divergence from the model system. Altered environments and other species can be identified as outliers from the model set. These results show that pattern recognition analysis has direct applicability to monitoring fermentation processes.     

Experimental observations consistent with a surface tension model of gliding motility of Myxococcus xanthus.

We have presented experimental evidence to support the model that gliding motility of Myxococcus xanthus is driven by surface tension. (i) Motility is inhibited by the addition of sufficient exogenous, nontoxic surfactants to swamp out the cells' own surfactant gradient. (ii) M. xanthus does not move polystyrene latex beads over its surface. (iii) Motility is prevented by elimination of an interfacial surface tension either by embedding the cells in soft agar or by placing them at an agar-aqueous interface. (iv) Wild-type cells excrete surfactant, whereas two nonmotile mutants excrete reduced amounts.     

Comparisons of mescal bean alkaloids with mescaline, delta 9-THC and other psychotogens

Prior to the discovery of peyote, many American Indian tribes of the Southwest apparently used seeds of the Texas Mountain Laurel. The seeds are commonly referred to as “mescal beans,” “red beans,” and “dry whiskey,” and were utilized by the Indians to induce visions and serve as a divinatory medium for various ceremonial purposes. Indications that this material is a part of the modern drug scene prompted studies in which the three major alkaloids from this plant (cytisine, sparteine, and methylcytisine) were compared with a variety of known psychoactive compounds [N, N-dimethyltryptamine (DMT), mescaline, psilocybin, amphetamine, Δ⁹ - tetrahydrocannabinol (THC), and pentobarbital] to determine whether similarities in behavioral effects exist. Rats were tested for gross locomotor activity, locomotor activity pattersn (“pauses” and “bursts”), locomotor skill (rotorod testing), and conditioned avoidance response. A Duncan's Multiple Range Comparison of all of the drugs at several dose levels of each revealed that the alkaloids produced responses similar to the responses produced by the known hallucinogenic drugs (mescaline, DMT, psilocybin) and clearly dissimilar to normal saline, amphetamine, pentobarbital, and THC.     

Comparative electrophoretic studies of crustacean neurosecretory hyperglycemic and melanophore-stimulating hormones from isolated sinus glands

Summary By disc electrophoresis of sinus gland extracts fromOrconectes limosus, Pacifastacus leniusculus andCarcinus maenas, marked differences in the electrophoretic mobility were demonstrated for the hyperglycemic neurohormone (HGH) from the 3 species. In the astacuran materials, 2 peaks of activity were observed whereas inCarcinus material only 1 peak could be demonstrated with certainty. Melanophore-stimulating activity (MDH) was resolved into several peaks, but 2 predominant peaks with almost identical mobility were observed in all 3 species. Electrophoresis of sinus gland material from the above species and fromHomarus gammarus, Procambarus clarkii, Astacus leptodactylus, A. astacus, Uca pugilator, Cancer magister, Eriocheir sinensis andHemigrapsus oregonensis revealed that all astacuran HGHs fall in one group of substances with low mobility whereas some of the brachyuran hormones (fromCarcinus, Uca andCancer) moved faster. The hormones ofEriocheir andHemigrapsus have mobilities similar to those of the astacuran hormones, but, like the other brachyuran hormones, they are not hyperglycemic in Astacura. HGH was found to be associated with the strongest staining band in the pherograms from all species investigated. It amounts to at least 10% of total sinus gland protein, that is about 0.5 g/gland inOrconectes and 0.6 g/gland inCarcinus. Accumulation of neurosecretory material in the neurohaemal organ is demonstrated by comparison of sinus gland and medulla terminalis extracts. In SDS gels, a large fraction of material (containing the HGH) in the mass range of 6000–8000 daltons is found in the the sinus gland whereas this fraction is barely demonstrable in the medulla terminalis. It was investigated whether cysteine-rich carrier proteins comparable to the vertebrate neurophysins exist in the sinus gland. No positive results were obtained either by aldehyde fuchsin staining of gels or by analysis of³⁵S-labeled sinus gland material. noreactivity in gels after electrophoresis of sinus gland extracts. No positive reaction was obtained with eluates from HGH containing slices. Recent amino acid analyses of HGH (Kleinholz, 1975; Keller and Wunderer, in preparation) do not suggest any similarities between crustacean HGH and vertebrate glucagon.     

Immunocytochemical identification of hyperglycemic hormone-producing cells in the eyestalk of Carcinus maenas

Summary Antiserum raised in rabbits against extracts of sinus glands from Carcinus and shown by several criteria to contain antibodies directed against the neurosecretory hyperglycemic hormone was used to locate the hormone-producing perikarya in the optic ganglia. By means of the double antibody fluorescence technique, selective staining of the large neurosecretory perikarya of the medulla terminalis ganglionic X-organ (MTGXO) and their axons is obtained. The axon endings of the sinus gland are also stained. None of the other groups of neurosecretory cells in the eyestalk shows fluorescence. Preabsorption of the antiserum with pure hyperglycemic hormone abolishes the fluorescence.Supported by the Deutsche Forschungsgemeinschaft (SFB 87, A 3; Ke 206/2). Thanks are due to E. Schmid (Ulm) for excellent technical assistance and to Prof. R. Martin and E. Weber for help and suggestions. A short version of parts of the results has been presented at theXth Conference of European Comparative Endocrinologists, Sorrento, May 1979     

Aspects of interspecific interaction betweenReithrodontomys megalotis andMicrotus montanus in one acre enclosures

1. Two one acre enclosures were cleared of all resident rodents, and then, one enclosure was seeded with founder populations of Reithrodontomys megalotis (grid M) and the other with Reithrodontomys megalotis plus Microtus montanus (grid I). Founder populations consisted of eight animals for each species introduced and a sex ratio of 1∶1.
2. Five parameters were measured for a period of one year. Data collection was started in September 1971, and ended in September 1972; enumeration was conducted twice a month for three days.
3. The five parameters measured were: (1) population density through time and individual growth rates; (2) reproduction; (3) survival of age and sex classes; (4) sex ratio; and (5) sizes of home ranges.
4. There were no significant differences in three out of the five parameters studied. Density estimates along with individual growth rates were not significantly different between the grids. Reproduction, including breeding season and efficiency of reproductive effort, showed no or very little variation due to interspecific interaction. Home range sizes did not appear to be significantly different between the grids. Survival of juvenile males on grid I seemed lower and juvenile males from grid I were significantly smaller although possibly younger than those of grid M. The sex ratio of grid I was significantly different from the expected 1∶1 ratio.
5. It is postulated that Reithrodontomys megalotis may regulate their density by alteration of their sex ratios.

     

The posterior opening of the pterygopalatine fossa and the position of the pterygopalatine ganglion

Our material consisted of 100 adult skulls and 50 adult half-head sections where the posterior opening of the pterygopalatinate fossa and the position of the pterygopalatinate ganglion were investigated. The report contains the following specifics: 1. Course and contents of the pterygoidal canal as well as the topographical relations of the canal to the sphenoid sinus. 2. Positioning of the axis of the foramen rotundum to the various planes of the head and its contents. 3. Course of the vomerovaginal canal and its contents. 4. Distances of the pterygopalatinate ganglion to important reference points of the head and to the median-sagittal line.     

Transition between isozymic forms of enolase during in vitro differentiation of neuroblastoma cells—II

An analysis of enolase expression during differentiation of neuroblastoma clones in homogeneous culture is presented. The enolases expressed in these neuroblast-like cells are identical to those of mouse brain with respect to the examined properties.Our biochemical investigation has premitted us to demonstrate formally that neuroblastoma cells undergo a transition from the embryonic αα form to the neuronal γγ form and contain both enolases as well as the αγ hybrid form during maturation. These results suggest that the same phenomenon must exist in vivo for neuroblasts. In neuroblastoma cells, an increase in both αγ and γγ neuron specific enolases is related to cell maturation and expression of the αγ form precedes that of the γγ form during differentiation. Modulation of neuronal enolase activities is similar in the various conditions of differentiation studied and appears not to be necessarily related with morphological differentiation, although concomitant with an arrest of cell division. The evolution of specific neuronal enolases in neuroblastoma cells parallels that observed in vivo, in brain from embryonic day 15 to post-natal day 7. Moreover, at least one treatment (dimethylsulfoxide) causes an important decrease in the high specific αα activity of these cells as occurs in vivo. This enolase can therefore also be considered as a biochemical marker for neuroblastoma maturation.As observed with other markers and other cell types, neuroblastoma cells in culture express an immature biochemical differentiation of the enolase isozymes.     

Histone interactions in solution and susceptibility to denaturation.

Histone interactions in solution may depend upon treatments used for purification. Optical rotatory dispersion and sedimentation-velocity measurements have been made in a reference solvent, before and after exposure to various treatments, to investigate histone susceptibility to irreversible denaturation. Some acid conditions and urea and guanidine solutions may denature. Interaction studies performed on nondenatured histones indicate that the dimer, (H4)(H3), and tetramer, (H4)2(H3)2, dissociate to monomers at low ionic strength. Sedimentation-velocity experiments suggest a model for the (H4)2(H3)2 tetramer, with a compact semispherical center and four protruding amino-terminal regions. Fractions H2a and H2b interact to form the mixed dimer in equilibrium with monomers. Fraction H2a self-associates readily to dimers, tetramers, and octamers, while fraction H1 associates only weakly to form dimers.