Message from the President of the Society

The circadian rhythm of locomotor activity of the field mouse Mus booduga was studied and single animal phase response curves (PRCs) (n = 8) were constructed for 15-min daylight pulses of 1000 lux intensity. The light pulses, presented at different phases of the circadian cycle, evoked advancing and delaying phase shifts (ΔPHs) depending on the circadian time (CT) of light pulse application. ΔPHs by light pulses applied at the same phase are strongly correlated with the animals' circadian period (τ). The results indicate a significant correlation between (i) τ and the area under the advance zone of the PRC (A) (r = +0.72, p > 0.05), (ii) τ and the area under the delay zone of the PRC (D) (r = -0.98, p > 0.00001), (iii) τ and the difference between the area under delay and advance zone of PRC (D-A) (r = -0.97, p > 0.00001), and (iv) between τ and ΔpHs (at various phases of the circadian cycle) and further suggest that the waveform and time course of PRC depend on the animals' endogenous period (τ). (Chronobiology International, 13(6), 401–409, 1996)     

New brachiopod species from the order acrotretida from the Cambrian of central Kazakhstan

Four new brachiopod species from the order Acrotretida (class Lingulata): Picnotreta saryarkensis sp. nov., Stilpnotreta propria sp. nov., Anabolotreta firma sp. nov., and Batenevotreta variabilis sp. nov. are described from the Agyrek Mountains, northeastern Central Kazakhstan from two olistoliths of limestones, which contain the uppermost Middle and lowermost Upper Cambrian fauna and are located in the Upper Ordovician olistostrome. These new species supplement the described earlier brachiopod assemblage from this region (Koneva and Ushatinskaya, 2008). Strong age and intraspecific variability of some Kazakh acrotretids is shown.     

Separation of the Mg-ATPases from the Ca-Phosphatase Activity of Microsomal Membranes Prepared from Barley Roots

Two methods for preparing membrane fractions from barley (Hordeum vulgare cv California Mariout 72) roots were compared in order to resolve reported differences between the characteristics of the plasma membrane ATPase of barley and that of other species. When microsomal membranes were prepared by a published procedure and applied to a continuous sucrose gradient, the membranes sedimented as a single broad band with a peak density of 1.16 grams per cubic centimeter (g/cm³). Activities of NADH cytochrome (Cyt) c reductase, Ca²⁺-ATPase, and Mg²⁺-ATPase were coincident and there was little ATP-dependent proton transport anywhere on the gradient. When the homogenization procedure was modified by increasing the pH of the buffer and the ratio of buffer to roots, the microsomal membranes separated as several components on a continuous sucrose gradient. A Ca²⁺-phosphatase was at the top of the gradient, NADH Cyt c reductase at 1.08 g/cm³, a peak of ATP-dependent proton transport at 1.09 to 1.12 g/cm³, a peak of nitrate-inhibited ATPase at 1.09 to 1.12 g/cm³, and of vanadate-inhibited ATPase at 1.16 g/cm³. The Ca²⁺-phosphatase had no preference for ATP over other nucleoside di- and tri-phosphates and was separated from the vanadate-inhibited ATPase on a sucrose gradient; approximately 70% of the Ca²⁺-phosphatase was removed from the microsomes by washing with 150 millimolar KCl. The vanadate-sensitive ATPase required Mg²⁺, was highly specific for ATP, and was not affected by the KCl wash. These results show that barley roots have a plasma membrane ATPase similar to that of other plant species.     

Formations of Oligopeptides from Dipeptides by the Protease from Streptomyces cellulosae

The protease from Streptomyces cellulosae formed more turbidity in a 16% soybean protein hydrolysate in the initial stage of the reaction than α-chymotrypsin did, when the proteolytic activity of the protease was same as that of α-chymotrypsin. In highly concentrated solutions (2.5%) of various dipeptides, oligopeptides were produced by condensation by the protease. The oligopeptides formed were (l-Leu-Gly)₂ and (l-Leu-Gly)₃ from l-Leu-Gly, (l-Phe-l-Val)₂ from l-Phe-l-Val, (l-Val-l-Phe)₂ and (l-Val-l-Phe)₃ from l-Val-l-Phe, and (l-Leu-l-Met)₂ and (l-Leu-l-Met)₃ from l-Leu-l-Met.     

Further Amphoricarpolides from the Surface Extracts of Amphoricarpos Complex from Montenegro

Analysis of composition of sesquiterpene lactone fraction of leaf cuticular neutral lipids of Amphoricarpos complex from two different localities in north Montenegro, i.e., canyon of river Tara (A. autariatus ssp. autariatus) and mountain Zeletin (A. autariatus ssp. bertisceus) afforded sesquiterpene lactones with guaianolide skeletons (17 compounds), so called amphoricarpolides, typical for this genus. Nine of them, 9 – 17 , were new compounds, and their structures were elucidated by detailed analyses of IR, NMR, and MS data.     

Cytotoxic metabolites from the cultures of endophytic fungi from Panax ginseng

Two strains of endophytic fungi, Penicillium melinii Yuan-25 and Penicillium janthinellum Yuan-27, with strong anti-Pyricularia oryzae activity, were obtained from the roots of Panax ginseng. Based on bioactivity-oriented isolation, a new benzaldehyde derivative, ginsenocin (1), together with six known compounds, methyl 2,4-dihydroxy-3,5,6-trimethylbenzoate (2), 3,4,5-trimethyl-1,2-benzenediol (3), penicillic acid (4), mannitol (5), ergosterol (6), and ergosterol peroxide (7), were separated from the EtOAc extract of Yuan-25 culture, while brefeldin A (8) was isolated as the major constituent from the EtOAc extract of Yuan-27 culture. The chemical structures were determined based on spectroscopic methods. All the isolated compounds 1–8 were evaluated for their cytotoxicity against six human cancer cell lines. Brefeldin A (8) was the most cytotoxic constituent against all the tested cell lines with IC₅₀ values <0.12 μg/ml, while ginsenocin (1) and penicillic acid (4) also exhibited potent cytotoxicity with IC₅₀ values ranging from 0.49 to 7.46 μg/ml. Our results suggest that endophytic fungi isolated from P. ginseng are a promising natural source of potential anticancer agents.     

Insights from the Genome Annotation of Elizabethkingia anophelis from the Malaria Vector Anopheles gambiae

Elizabethkingia anophelis is a dominant bacterial species in the gut ecosystem of the malaria vector mosquito Anopheles gambiae. We recently sequenced the genomes of two strains of E. anophelis, R26^T and Ag1, isolated from different strains of A. gambiae. The two bacterial strains are identical with a few exceptions. Phylogenetically, Elizabethkingia is closer to Chryseobacterium and Riemerella than to Flavobacterium. In line with other Bacteroidetes known to utilize various polymers in their ecological niches, the E. anophelis genome contains numerous TonB dependent transporters with various substrate specificities. In addition, several genes belonging to the polysaccharide utilization system and the glycoside hydrolase family were identified that could potentially be of benefit for the mosquito carbohydrate metabolism. In agreement with previous reports of broad antibiotic resistance in E. anophelis, a large number of genes encoding efflux pumps and β-lactamases are present in the genome. The component genes of resistance-nodulation-division type efflux pumps were found to be syntenic and conserved in different taxa of Bacteroidetes. The bacterium also displays hemolytic activity and encodes several hemolysins that may participate in the digestion of erythrocytes in the mosquito gut. At the same time, the OxyR regulon and antioxidant genes could provide defense against the oxidative stress that is associated with blood digestion. The genome annotation and comparative genomic analysis revealed functional characteristics associated with the symbiotic relationship with the mosquito host.     

Methylation studies of the polysaccharides resulting from sequential Smith-degradations of the galactan from the snail strophocheilus oblongus

When the galactan from the albumen glands of the snail Strophocheilus oblongus was submitted to three Smith-degradations, the degraded polysaccharide, isolated in 6% yield, was much more linear. Methylation analysis showed that the Smith-degraded polysaccharide gave an increased percentage of 2,4,6-tri-, decreased percentages of 2,3,4,6-tetra- and 2,4-di-, and a large variation in the amount of 2,3,4-tri-O-methyl-d-galactose. The sugars in the polysaccharide which result in the formation of 2,3,4,6-tetra- and 2,3,4-tri-O-methyl-d-galactose are destroyed in subsequent degradation procedures. The above observations suggest that the degradation by periodate oxidation proceeds via non-reducing end-groups and through some internal residues that are exposed as the degradation proceeds. As a result of the overall process, new non-reducing end-groups are formed and new (1 → 6)-linked d-galactose residues are then exposed. The isolation of glycosides of low molecular weight supports the suggestion that the molecule, in the main, is sequentially degraded from the external layers.     

Relatedness of the flagellins from methanogens

Purified flagellar filaments isolated from six methanogens were composed of multiple flagellins. Two flagellins were present in Methanococcus deltae (M _r =34000 and 32000), Methanoculleus marisnigri (M _r =31000 and 25500) and Methanococcus jannaschii (M _r =31000 and 27500), three in Methanothermus fervidus (M _r =34000, 25000 and 24000) and four or more in both Methanococcus vanniellii and Methanococcus maripaludis (M _r ranging from 27500 to 32000). The flagellins of M. fervidus and M. deltae reacted positively with glycoprotein-specific stains. The flagellins of M. deltae, M. maripaludis and M. vannielii were closely related to those of M. voltae based on cross-reactivity with antisera raised against M. voltae flagellins and homology with flagellin-specific oligonucleotide probes to the N-terminus and leader peptide of M. voltae flagellins. Similarities appear to exist among the flagellins of M. fervidus, M. marisnigri and Halobacterium halobium based on cross-reactivity with antisera produced against the flagella of Methanospirillum hungatei JF1. The N-termini of the flagellins from the mesophilic Methanococcus spp. and M. marisnigri show homology with the N-termini of other archaebacterial flagellins. These N-termini may undergo a modification involving removal of a leader peptide.     

Antibacterial metabolites from Australian macrofungi from the genus Cortinarius

In this study, ethyl acetate and aqueous fractions from 117 collections of Australian macrofungi belonging to the mushroom genus Cortinarius were screened for antimicrobial activity against Staphylococcus aureus and Pseudomonas aeruginosa. Overall, the lipophilic fractions were more active than the aqueous fractions. The ethyl acetate fractions of most or all collections of 13 species, namely Cortinarius ardesiacus, C. archeri, C. austrosaginus, C. austrovenetus, C. austroviolaceus, C. coelopus, C. [Dermocybe canaria]², C. clelandii, C. [D. kula], C. memoria-annae, C. persplendidus, C. sinapicolor, C. vinosipes and forty seven collections of un-described Cortinarius species exhibited IC₅₀ values of ?0.09 mg/mL against S. aureus. In contrast, most or all collections of only four species, namely C. abnormis, C. austroalbidus, C. [D. kula], C. persplendidus, and eleven un-described Cortinarius collections exhibited similar effects against P. aeruginosa (IC₅₀ ? 0.09 mg/mL). Anthraquinonoid pigments isolated from C. basirubescens together with emodin physcion and erythrogluacin were assessed for their antimicrobial activity. The fungal octaketides austrocortilutein, austrocortirubin, torosachrysone, physcion and emodin were found to strongly inhibit the growth of S. aureus (IC₅₀ 0.7–12 μg/mL) whereas only physcion and emodin exhibited potency against P. aeruginosa (IC₅₀ 1.5 and 2.0 μg/mL, respectively).     

Cephalopod and brachiopod fossils from the Pacific: Evidence from the Upper Cretaceous of the Magellan Seamounts

Maastrichtian cephalopods and a brachiopod were dredged from the Butakov, Fedorov, Kotsebu, Il’ichev, Govorov, Gelendzhik, and Ita-Mai-Tai guyots in the Magellan Seamounts. The ammonoids Hypophylloceras sp., Phyllopachiceras sp., Anagaudryceras? sp. A, Anagaudryceras? sp. B, Gaudryceras aff. propemite Marshall, Gaudryceras sp., and Pseudophyllites cf. indra (Forbes), and the single brachiopod Basiliolidae gen. and sp. indet. are a first discovery in this oceanic region, following earlier finds of belemnites (Dimitobelus? sp., Dimitobelidae gen. and sp. nov., and Belemnitella? sp.), and two ammonoid species (Zelandites aff. japonicus Matsumoto and Tetragonitidae gen. and sp. indet.). The Late Cretaceous Magellan Seamounts dimitobelid belemnite fauna shows affinities with southern high latitude forms (New Zealand) and the ammonoid fauna with northern, middle and high latitude ones (Hokkaido-Sakhalin and/or Kamchatka). This suggests by the end of the Cretaceous major surface palaeo-currents from S and N sides in direction of the central Palaeo-Pacific, a position coinciding with the plate tectonic reconstruction of the Magellan Seamounts.     

Chemical Composition of Essential Oil from Mosses from the Brazilian Atlantic Forest

This study aimed to report the unprecedented volatile composition of the mosses Phyllogonium viride BRID, Orthotichella rigida (MÜLL.HAL.) B. H. ALLEN & MAGILL and Schlotheimia rugifolia (HOOK.) SCHWÄGR occurring in the Brazilian Atlantic Forest, in order to elucidate the chemical composition of these species and enrich the chemotaxonomic knowledge of mosses. 28 compounds were identified, the major constituent being hexadecanoic acid, also known as palmitic acid, specifically P. viride com (38.55 %), O. rigida com (17.17 %) and S. rugifolia com (24.94 %), followed by phytol, P. viride com (3.92 %), O. rigida com (28.57 %) and S. rugifolia com (36.13 %). In addition, there was a prevalence of aliphatic hydrocarbons (25 %) and fatty acids (17.8 %) in the evaluated samples. These data contribute to the generation of new scientific information about the chemical constitution of mosses, still little studied, enriching the chemotaxonomic collection of the taxon.     

Items from the Literature

LIPP, WALTHER. Histochemische Methoden. Twenty-four pages per issue; 3 issues (No. 12, 13 and 14) during 1957. R. Oldenbourg, Miinchen 1, Germany. DM 30-for 6 issues; DM 6—per single copy. (In German)

STACH, E. Auflicht-Mikroslcopie mit Immersions-Kappen. Mikroskopie, 12, 232-42. 1957.

JOHNSON, C. Time savers in the paraffin technique. Turtox News, 36, 126-7. 1958.

JONSSON, N., and LAGERSTEDT, S. Losses of nucleic acid derivatives from fixed tissues during flattening of paraffin sections on water. Experientia, 14, 157-9. 1958.

SCHAUMKELL, K. W., STANGE, H. H., and DÖRFFLER, P. Methodische Untersuchungen über die pH-abhängige Aufname saurer und basischer Hellfeld-Diachrome in der Nebennierenrinde. Mikroskopie, 12, 227-32. 1957.

SCHEFTHALER, MARIA, and MAYET, A. Eine Modification der Bielschowsky Methode für periphere Nervenfasern. Mikroskopie, 12, 298-801. 1958.

LANGENBUCH, R. Beitrag zur Färbung von Einschluszkörpern (Polyedern) in Blut- und Gewebeausstrichen viruskranker Insecten. Mikroskopie, 12, 267-8. 1957.

SHUGAR, D., and BARANOWSKA, J. Quantitative Gram staining with labelled iodine. Nature, 181, 357-8. 1958.

ARVY, L., and GABE, M. Comparison des activitiés aliestérasique mises en évidence par les techniques au tween et a l'acétate de naphthyle. Ann. d'Histochimie, 2, 141-8. 1957.

VERNE, J., and WEGMANN, R. Problèmes actuels de la cytochimie des estérases (carboxyl estérases and cholinestérases). Ann. d'Histochimie, 2, 75-107. 1957.

WEGMANN, R. Les carbonyles lipidiques. III. Les facteurs associés au prétraitement. Ann. d'Histochimie, 2, 197-214. 1957.     

Phylogeography of the mallard Anas platyrhynchos from Eurasia inferred from sequencing of the mtDNA control region

Phylogenetic relationships, demographic history, and geographic distribution of the mtDNA haplotypes of the mallard Anas platyrhynchos were examined in three populations, Indian, Northern European, and Far Eastern. Two divergent halotype groups, A and B, were found in the Far Eastern population, while haplotypes identified in Northern European and Far Eastern populations were exclusively of the A group. The presence of B group haplotypes in the Far Eastern population can be explained either in terms of hybridization of the mallard with spot-billed duck Anas zonorhyncha at the south of the Russian Far East, or by the mtDNA paraphyly in mallards. In general, mallards from Eurasia were characterized by low genetic population differentiation along with slightly expressed phylogeographic structure. The most differentiated was the population from India (??_st = 0.076?0.077), while the difference between Northern European and Far Eastern populations was extremely low (??_st = 0.0029). Differentiation of Anas platyrhynchos Indian population was determined by the fact that a part of the population, inhabiting southern and eastern coasts of the Hindustan Peninsula, was resident.     

Purification and Characterization of Proteins from the 2S Fraction from Seeds of the Brassicaceae Family

The 2S protein fractions from Brassica rapa, Brassica oleracea,and Brassica napus seeds have been obtained and their componentspurified and characterized. These albumins represent about 13%of the total seed protein extracted with saline buffer. Thecalculated molecular weights of the eleven purified proteinsare very similar, about 14 500 Daltons, although two componentsisolated from B. napus exhibit a lower molecular weight. Theamino acid compositions of the isolated proteins present a seriesof common features: a high content of Cys and basic residuesas well as a very high amount of Pro (15%) and Glx (30%). Theeleven purified proteins cross-react with antibodies againstSin a I, the 2S protein from yellow mustard seeds. The obtainedresults suggest the existence of homology between the 2S albuminsof Brassicaceae seeds. Key words: Brassicaceae, seeds, storage protein     

Investigating the Allosteric Regulation of YfiN from Pseudomonas aeruginosa: Clues from the Structure of the Catalytic Domain

Pseudomonas aeruginosa is responsible for a plethora of biofilm mediated chronic infections among which cystic fibrosis pneumonia is the most frightening. The long-term survival strategy of P. aeruginosa in the patients lungs is based on a fine balance of virulence vs dormant states and on genetic adaptation, in order to select persistent phenotypes as the small colony variants (SCVs), which strongly correlate with antibiotic resistance and poor lung function. Recent studies have coupled SCV with increased levels of the signaling molecule cyclic di-GMP, and demonstrated the central role of the diguanylate cyclase YfiN, part of the tripartite signaling module YifBNR, in c-di-GMP dependent SCV regulation. YfiN, also called TpbB, is a multi-domain membrane enzyme connecting periplasmic stimuli to cytosolic c-di-GMP production by an allosteric inside-out signaling mechanism that, due to the lack of structural data, is still largely hypothetical. We have solved the crystal structure of the catalytic domain (GGDEF), and measured the enzymatic activity of the cytosolic portion in real-time by means of a newly developed method. Based on these results we demonstrate that, unlike other diguanylate cyclase, YfiN does not undergo product feedback inhibition, and that the presence of the HAMP domain is required for dimerization and catalysis. Coupling our structural and kinetic data with an in silico study we are now able to propose a model for the allosteric regulation of YfiN.     

Identification of Janthinobacterium lividum from the soils of the islands of Scotia Ridge and from Antarctic peninsula

Summary Eight isolates of bacteria from the soils of maritime Antarctica and Antarctic peninsula have been identified as members of the genus Janthinobacterium. Based on their morphology, physiological characteristics, biochemical characteristics and mole percent G+C content of their DNA six of them have been identified as J. lividum and the remaining two as atypical J. lividum. The Antarctic J. lividum unlike the mesophilic type strains were unique in that they could grow at pH 4, could produce acid from trehalose and none of them could tolerate more than 2.9% NaCl.     

Onward from the cradle

This essay records a voyage of discovery from the “cradle of cell biology” to the present, focused on the biology of the oldest known cell organelle, the cilium. In the “romper room” of cilia and microtubule (MT) biology, the sliding MT hypothesis of ciliary motility was born. From the “summer of love,” students and colleagues joined the journey to test switch-point mechanisms of motility. In the new century, interest in nonmotile (primary) cilia, never lost from the cradle, was rekindled, leading to discoveries relating ciliogenesis to autophagy and hypotheses of how molecules cross ciliary necklace barriers for cell signaling.     

Items from the Literature

Conn, H. J. Biological Stains. 7th ed., 355 pages, 27 figures, 9 × 6 inches, hard covers. The Williams &;: Wilkins Company, Baltimore 2, Maryland. $9.00. 1961.

Electron Microscopy Reimer, L., and GADACZ, H. Zur Schichtdickenkontrolle bei der elektronenmikroskopischen Schrägbeschattungsmethode. Z. wiss. Mikr., 65, 105-17. 1961.

Dyes and Their Biological Uses Macconaill, M. A., and GURR, E. The faviolic acid group of stains in histology. Irish J. Med. Sci., 1962, 1-12. 1962.

Animal Microtechnic Berres, H. H. Zur Darstellung peripherer Nervenfasem in der Haut. Z. wiss. Mikr., 65, 63-9. 1961.

Elias H., Henning A., and ELIAS P. M. Contributions to the geometry of sectioning. V. Some methods for the study of kidney structure. Z. wiss. Mikr., 65, 70-82. 1961.

Ewen, A. B. An improved aldehyde-fuchsin staining technique for neurosecretary products in insects. Tr. Amer. Micr. Soc, 81, 94-6. 1962.

Movat, Henry Z., STEINER, JAN W., and HUHN, DIETER. The fine structure of the glomerulus in acute glomerulonephritis. Lab. Investig., 11, 117-35. 1962.

Shaver, Evelyn L. The chromosomes of the opossum, Didelphis virginiana. Canad. J. Genet. Cytol. 4, 62-8. 1962.

Microorganisms Laird, Marshall. Trichodinids and other parasitic protozoa from the intertidal zone at Nanaimo, Vancouver Island. Canad. J. Zool., 39, 833-44. 1961.

Histochemistry Burstone, M. S. New histochemical techniques for the demonstration of tissue oxidase (cytochrome oxidase). I. Hisiochem. Cytochem., 7, 112-22. 1959.

Glenner, G. G., and LILLIE, R. D. Observations on the diazotization-coupling reaction for the histochemical demonstration of tyrosine: metal chelation and formazan variants. J. Histochem. Cytochem., 7, 416-22. 1959.

Hess, R., and Pearse, A. G. E. Histochemical demonstration of uridine diphosphate glucose dehydrogenase. Experientia, 15, 317-8. 1961.

LOVE, R., and LILES, R. H. Differentiation of nucleoproteins by inactivation of protein-bound amino groups and staining with toluidine blue and ammonium molybdate. J. Histochem. Cytochem., 7, 164-81. 1959.

Zimmerman, H., and Pearse, A. G. E. Limitations in the histochemical demonstration of pyridine nucleotide-linked dehydrogenases (“nothing dehydrogenase”). J. Histochem. Cytochem. 7, 271-5. 1959.     

Insight from the structural molecular model of cytidylate kinase from Mycobacterium tuberculosis

Mycobacterium tuberculosis is a gram-positive bacterium causes tuberculosis in human. H37Rv strain is a pathogenic strain utilized for tuberculosis research. The cytidylate mono-phosphate (CMP) kinase of Mycobacterium tuberculosis belongs to the family nucleoside mono-phosphate kinase (NMK), this enzyme is required for the bacterial growth. Therefore, it is important to study the structural and functional features of this enzyme in the control of the disease. Hence, we developed the structural molecular model of the CMP kinase protein from Mycobacterium tuberculosis by homology modeling using the software MODELLER (9v10). Based on sequence similarity with protein of known structure (template) of Mycobacterium smegmatis (PDB ID: 3R20) was chosen from protein databank (PDB) by using BLASTp. The energy of constructed models was minimized and the qualities of the models were evaluated by PROCHECK and VERRIFY-3D. Resulted Ramachandran plot analysis showed that conformations for 100.00% of amino acids residues are within the most favored regions. A possible homologous deep cleft active site was identified in the Model using CASTp program. Amino acid composition and polarity of that protein was observed by CLC-Protein Workbench tool. Expasy''s Prot-param server and CYC_REC tool were used for physiochemical and functional characterization of the protein. Studied of secondary structure of that protein was carried out by computational program, ProFunc. The structure is finally submitted in Protein Model Database. The predicted model permits initial inferences about the unexplored 3D structure of the CMP kinase and may be promote in relational designing of molecules for structure-function studies.