The 108-kDA peptidylglycine alpha-amidating monooxygenase precursor contains two separable enzymatic activities involved in peptide amidation

A 43-kDa protein factor that increases the ability of purified bovine peptidylglycine alpha-amidating monooxygenase (PAM)-A and -B to produce alpha-amidated peptides at physiological pH was purified to homogeneity from bovine neurointermediate pituitary. At each step of the purification, the amount of activity correlated with the amount of protein detected on Western blots by antibody to bovine PAM(561-579). In the bovine neurointermediate pituitary the 108-kDa PAM precursor protein is cleaved to form a peptidylglycine alpha-hydroxylating monooxygenase and a peptidyl-alpha-hydroxyglycine alpha-amidating lyase, which function sequentially in the 2-step formation of alpha-amidated peptides.     

Behavioral changes of a juvenile gorilla after a transfer to a more naturalistic environment

Play behavior and stress-associated behavior of a captive juvenile gorilla were observed before and after his transfer to a larger and more natural environment. The gorilla was observed for 4 months after the transfer and at 1 and 4 years after the transfer. Throughout his first month in the new environment play decreased dramatically. Although play subsequently increased again 2 months and 1 year after the transfer, it never reached the levels of play in the old environment. Four years after the move his play had decreased again to the low level of his first month in the new environment. Two of his stress-associated behaviors, coprophagy and regurgitation/reingestion, decreased after the transfer. Self-clasping behavior increased initially in the new environment and remained at high levels 1 year after the move. Four years after the move his self-clasping behavior was significantly less than at 1 year after the move; however, it continued to be significantly greater than in the old environment. These findings suggest that larger and more natural environments do not necessarily result in more play activity or a reduction in all stress-associated behavior.     

Structure of the precursor to an enzyme mediating COOH-terminal amidation in peptide biosynthesis

Many bioactive peptides terminate with an amino acid alpha-amide at their COOH terminus. The enzyme responsible for this essential posttranslational modification is known as peptidyl-glycine alpha-amidating monooxygenase or PAM. We identified cDNAs encoding the enzyme by using antibodies to screen a bovine intermediate pituitary lambda gt11 expression library. Antibodies to a beta-galactosidase/PAM fusion protein removed PAM activity from bovine pituitary homogenates. The 108,207 dalton protein predicted by the complete cDNA is approximately twice the size of purified PAM. An NH2-terminal signal sequence and short propeptide precede the NH2 terminus of purified PAM. The sequences of several PAM cyanogen bromide peptides were localized in the NH2-terminal half of the predicted protein. The cDNA encodes an additional 430 amino acid intragranular domain followed by a putative membrane spanning domain and a hydrophilic cytoplasmic domain. The forms of PAM purified from bovine neurointermediate pituitary may be generated by endoproteolytic cleavage at a subset of the 10 pairs of basic amino acids in the precursor. High levels of PAM mRNA were found in bovine pituitary and cerebral cortex. In corticotropic tumor cells, levels of PAM mRNA and pro-ACTH/endorphin mRNA were regulated in parallel by glucocorticoids and CRF.     

A catalogue of some ecuadorean fermented beverages,with notes on their microflora

Summary A catalogue of indigenous fermented beverages produced by different ethnic groups in Ecuador has been compiled and the microflora of selected examples examined. A diversity of fermentation substrates was encountered depending on the climatic zone. The fermentations are typicallyLactobacillus spp.—yeast fermentations except for one which includes a mould fermentation by a mixed starter ofMoniha sitophila, Rhizopus stolonifer and aFusarium sp. A discussion is made of the role of these beverages in the human ecology of certain regions.

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Resumen Se ha confeccionado un catálogo de bebidas indígenas ecuatorianas producidas por distintos grupos étnicos, examinándose la microflora de algunos ejemplos seleccionados. Las fermentaciones son generalmente del tipoLactobacillus sp.—levaduras, excepto en un caso que incluye una fermentación fúngica iniciada de forma mixta porM. sitophila, R. stolonifer y unFusarium sp. Se discute el papel de estas bebidas en la ecologia humana de ciertas regiones.

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Résumé Un catalogue des boissons fermentées indigènes produites par divers groupes ethniques de l'Equateur a été compilé et les micro-flores des exemples sélectionnés ont été éxaminés. Les substrats de fermentation varient d'une région climatique à l'autre. Les fermentations sont généralement du typeLactobacillus sp — levures, sauf dans un cas qui comporte une fermentation par des moisissures, avec un mélange initial deMoniha sitophila, Rhizopus stolonifer et une espèce deFusarium. Le rôle de ces boissons dans l'écologie humaine de certaines régions est discuté.

     

Competition in the gradostat: the role of the communication rate

In this paper we study a mathematical model of competition between two species of microorganisms for a single limiting nutrient in a laboratory device called a gradostat. A gradostat consists of several (we consider only two) chemostats (CSTR's) connected together so that material can flow between the vessels in such a way that a nutrient gradient is established. Our model is a slightly modified version of one considered recently by Jäger et al. [3], in that the rate of exchange of material between the two vessels (the communication rate) is allowed to differ from the dilution rate. The outcome of competition turns out to be surprisingly sensitive to variation of the communication rate. We identify several coexistence regimes in parameter space and describe a method for obtaining operating diagrams for given pairs of competing microorganisms.Research supported in part by NSF Grant DMS 8521605     

Carbohydrate constituents of Lessonia trabeculata

D-mannitol was the only low-molecular weight carbohydrate isolated from ethanolic extracts of Lessonia trabeculata blades. After sequential extraction with water, acid and alkali, laminaran, fucose-containing polysaccharides and alginic acid were also isolated. Fucose-containing polysaccharide from the acidic extract was separated into three fractions by ion exchange chromatography. Alginic acid was the major polysaccharide obtained in the sequential extraction.     

C1q,a collagen-like complement subcomponent,in dermatosparactic cattle: its extracellular modification is not affected by lack of procollagen N-terminal proteinase (pN-proteinase)

C1q, a collagen-like complement protein, was purified from the serum of a ddermatosparactic calf which lacks procollagen N-terminal proteinase (pN-proteinase). The specific hemolytic activity of the serum Clq from the dermatosparactic animal was identical to that of C1q from a normal calf. Gel-filtration of serum from dermatosparactic calf, on Sepharose 6B, showed the presence of C1q-antigenic material at only one position which was identical to the elution position of normal bovine C1q. No differdence, under dissociating conditions, could be seen in the size of the chains of C1q in specific immunoprecipitates isolated from the sera of dermatosparactic and normal animals, as judged by polyacrylamidegel electrophoresis (PAGE) in the presence of sodium dodecyl sulfate (SDS). The C1q from the dermatosparactic animal showed the same N-terminal amino acid and typtic-digest peptide pattern on HPLC as C1q from the normal calf. These results strongly suggest that pN-proteinase is not involved in the extracellular processing of C1q.     

MICROTUBULES ASSOCIATED WITH SIMULTANEOUS CYTOKINESIS OF COENOCYTIC MICROSPOROCYTES

Microtubule arrays associated with simultaneous cytokinesis in the coenocytic microsporocytes of Lonicera japonica and Impatiens sultani were studied by indirect immunofluorescence. The future division planes are not predicted prior to meiosis by either a preprophase band of microtubules or cytoplasmic lobing. Cleavage planes appear to be determined by position of the four haploid nuclei and the development of postmeiotic microtubule systems. Perpendicular second division spindles in Lonicera result in tetrahedrally arranged tetrads while parallel spindles in Impatiens result in tetragonal arrangement. Immediately following meiosis bands of microtubules, the secondary spindles, develop between both sister and nonsister nuclei. These arrays give way to systems of microtubules that radiate equally from each of the four nuclei in the coenocytic sporocyte. Simultaneous cytokinesis is initiated by centripetal wall deposition at the periphery of the sporocyte and proceeds along planes marked by interaction of the opposing arrays of nuclear-based microtubules.