Transformation of Atriplex gmelini plants from callus lines using Agrobacterium tumefaciens

Atriplex gmelini plants were regenerated via organogensis from hypocotyl explants. Callus lines were induced from the hypocotyl explants on Linsmaier and Skoog (LS) medium supplemented with 1 M benzyladenine and 5 M -naphthaleneacetic acid in the dark. Shoots were regenerated from the callus lines on LS medium supplemented with 20 M thidiazuron and 0.1 M -naphthaleneacetic acid under a high-intensity light condition (450 mol m^–2 s^–1). The regenerated shoots were rooted on LS medium without growth regulators to obtain fully developed plants. We succeeded in transforming Atriplex gmelini from callus lines using Agrobacterium tumefaciens.     

Cloning bacterial genes with plasmid λdv

Summary Fragments ofEscherichia coli DNA carrying genes for -galactosidase, or for biosynthesis of guanine or biotin were recombined in vitro with dv DNA. The cloned recombinant molecules recovered from transformedE. coli cells consisted of a biologically functional bacterial DNA fragment and, except for dv-bio30-7, two dv monomer units: one of the dv units was used as the insertion site for the bacterial DNA, whereas the other was intact, and seemed to be responsible for the replication of the recombinant plasmid. The process which gives rise to these recombinant molecules at high frequency from mixtures of monomeric dv DNA's and bacterial DNA fragments is discussed.     

Sarcocystis inghami n. sp. (Sporozoa: Sarcocystidae) from the skeletal muscles of the Virginia opossum Didelphis virginiana in Michigan

This report describes the newly identified Sarcocystis inghami n. sp. from the skeletal muscles of opossums (Mammalia: Didelphidae) that were collected from south central Michigan (42° 43-42° 79N, 84° 18-84°mathtype="display">6'W), USA. The new species is distinguished from all species described from North and South American opossums by the distinctive morphology of the villar protrusions on the cyst wall. Sarcocysts of S. inghami are microscopic, up to 700 m long and 110 m wide. The sarcocyst wall is up to 7 m thick, with long, stalked protrusions which average 5.5 × 1.2 m. These are constricted at the base, expanded laterally, rounded off distally and occasionally bifid. The villar protrusions have numerous microtubules without electron–dense bodies that extend from the tips into the granular layer. Bradyzoites are 10.7 × 4.3 (8––12 × 4––5) m. This is the second species of Sarcocystis sarcocyst described from the Virginia opossum in North America.     

Linkage mapping of the cystathionine β-synthase (CBS) gene on human chromosome 21 using a DNA polymorphism in the 3′ untranslated region

We used single-strand conformation polymorphism (SSCP) to detect DNA polymorphisms in the 3 untranslated (3UT) region of the gene for cystathionine -synthase (CBS). A polymorphism due to a T-to-C substitution at nucleotide 549 of the 3UT region with heterozygosity of 46% has been identified. Genotypes for this polymorphism have been obtained in all of the informative CEPH families, and CBS has been placed in the linkage map of human chromosome 21.     

Cystic kidneys

Summary According to the classification of Osathanondh and Potter of cystic kidneys we give an overview of the different types of cystic changes taking genetic aspects into account. Usually pathoanatomic types do not represent genetic entities: All type I kidneys are transmitted in an autosomal recessive way with varying clinical symptoms; in rare cases they even present in adults. The relationship to congenital hepatic fibrosis, cystic liver, and to the Caroli syndrome is discussed. Type II kidneys are usually not genetic in origin; but they may occur as part of several syndromes. Rarely genetic factors might contribute to type II kidneys that may present as familial cases of Potter syndrome (renal non-function syndrome). Type IV kidneys, although different in their pathoanatomic picture can be regarded according to a common pathogenetic theory as part of the spectrum of malformations as in type II. Therefore the genetic interpretation of type II kidneys also applies to type IV lesions. Type III kidneys include autosomal dominant polycystic kidney disease. This type may already present in childhood; the first prenatal diagnosis by ultrasonography is described in detail. Furthermore type III changes are part of syndromes or non-hereditary malformation complexes, and often present only as mild manifestations. Diseases with isolated involvement of the medulla (juvenile nephronophthisis/medullary cystic disease) or cortex are described as part of the differential diagnosis, they are heterogeneous and genetically only partly understood. Syndromes with cystic kidneys are reviewed as well as the possibilities of prenatal diagnosis of cystic diseases. Reliable prenatal diagnosis is only possible in type II, and possible in some of the other types. The nosology is improved if genetic information is taken into account.     

Adoptions expérimentales de larves entre des fourmis de genres différents (II):Myrmica laevinodis Nylander etAnergates atratulus Schenck

Résumé Nous avons fait élever des larves d'Anergates atratulus par des ouvrières deMyrmica laevinodis à 22°C. Pour y parvenir, il n'est pas utile de faire hivernerensemble les larves d'Anergates et les ouvrières deMyrmica. La présence de larves autochtones n'empêche pas lesMyrmica d'élever des larves d'Anergates. Dans toutes les expériences lesMyrmica ont été soumises au fridavant de recevoir des larves d'Anergates. Aucune reine deMyrmica n'a été utilisée dans ces expériences.Sur les 64 larves d'Anergates que nous avons utilisées, 38 se sont transformées en imagos. C'est au début de l'adoption et au moment des métamorphoses que périrent la plupart des 26Anergates perdus. Les femelles vécurent en général 2 ou 3 jours et cherchèrent très tôt à quitter le nid natal. Les mâles vécurent 2 à 3 semaines.

---

Summary Larvae ofAnergates atratulus were experimentally reared by workers ofMyrmica laevinodis, at 22°C. An overwintering of both larvae ofAnergates and workers ofMyrmica is not necessary for the success of that experiment. The presence of larvae ofMyrmica does not keep theMyrmica from rearing larvae ofAnergates. The workers ofMyrmica have been cooled, in all the experiments, before receiving larvae ofAnergates. No queen ofMyrmica have been used in that experiments.38 of the 64 larvae ofAnergates used became imagos. Most of the 26 lostAnergates died at the beginning of the adoption and during the metamorphosis. The females lived generally 2 or 3 days and tried, very early, to leave their native nest. The males lived 2 or 3 weeks.

---

Anergates atratulus Myrmica laevinodis, 22 . bmecme Anergates Myrmica. Myrmica Anergates. Myrmica Anergates. Myrmica . 64 Anergates , 38 . 26 Anergates 2 3 . 2 3 .

     

Genomic characterization of thermophilic Geobacillus species isolated from the deepest sea mud of the Mariana Trench

The thermophilic strains HTA426 and HTA462 isolated from the Mariana Trench were identified as Geobacillus kaustophilus and G. stearothermophilus, respectively, based on physiologic and phylogenetic analyses using 16S rDNA sequences and DNA–DNA relatedness. The genome size of HTA426 and HTA462 was estimated at 3.23–3.49 Mb and 3.7–4.49 Mb, respectively. The nucleotide sequences of three independent -phage inserts of G. stearothermophilus HTA462 have been determined. The organization of protein coding sequences (CDSs) in the two -phage inserts was found to differ from that in the contigs corresponding to each insert assembled by the shotgun clones of the G. kaustophilus HTA426 genome, although the CDS organization in another insert is identical to that in the HTA426 genome.     

Interaction of Amyloid Beta-Protein with Anionic Phospholipids: Possible Involvement of Lys28 and C-Terminus Aliphatic Amino Acids

Fibrillar amyloid beta-protein (A) is the major protein of amyloid plaques in the brains of patients with Alzheimer's disease (AD). The mechanism by which normally produced soluble A gets fibrillized in AD is not clear. We studied the effect of neutral, zwitterionic, and anionic lipids on the fibrillization of A 1-40. We report here that acidic phospholipids such as phosphatidic acid, phosphatidylserine, phosphatidylinositol (PI), PI 4-phosphate, PI 4,5-P₂ and cardiolipin can increase the fibrillization of A, while the neutral lipids (diacylglycerol, cholesterol, cerebrosides), zwitterionic lipids (phosphatidylcholine, phosphatidylethanolamine, sphingomyelin) and anionic lipids lacking phosphate groups (sulfatides, gangliosides) do not affect A fibrillization. A was found to increase the fluorescence of 1-acyl-2-[12-[ (7-nitro-2-1, 3-benzoxadiazol-4-yl) amino] dodecanoyl]-sn-glycero-3-phosphate (NBD-PA) in a concentration-dependent manner, while no change was observed with 1-acyl-2-[12-[(7-nitro-2-1, 3-benzoxadiazol-4-yl) amino] dodecanoyl]-sn-glycero-3-phosphoethanolamine (NBD-PE). Under similar conditions, other proteins such as apolipoprotein E, gelsolin and polyglutamic acid did not interact with NBD-PA. The order of interaction of amyloid -peptides with NBD-PA was A 1-43 = A 1-42 = A 17-42 > A 1-40 = A 17-40. Other A peptides such as A 1-11, A 1-16, A 1-28, A 1-38, A 12-28, A 22-35, A 25-35, and A 31-35 did not increase the NBD-PA fluorescence. These results suggest that phosphate groups, fatty acids, and aliphatic amino acids at the C-terminus end of A 1-40/A 1-42 are essential for the interaction of A with anionic phospholipids, while hydrophilic A segment from 1-16 amino acids does not participate in this interaction. Since positively charged amino acids in A are necessary for the interaction with negatively charged phosphate groups of phospholipids, it is suggested that Lys²⁸ of A may provide anchor for the phosphate groups of lipids, while aliphatic amino acids (Val-Val-Ile-Ala) at the C-terminus of A interact with fatty acids of phospholipids.     

Search for A-factor-Dependent Variants in Actinomycete Populations

A study of 28 nocardia-like, asporogenous, and oligosporous spontaneous morphological variants belonging to 23 species of streptomycetes revealed five strains producing regulators of the A-factor group. Streptomyces griseus 1439, which forms aerial mycelium and spores only in the presence of exogenous A-factor, was used as the test strain. Among the 28 spontaneous variants, three new A-factor-dependent strains were revealed, which represented the species Streptomyces griseus, S. citreofluorescens, and S. viridovulgaris subsp. albomarinus. These weakly differentiated variants did not produce A-factor and behaved as its recipients, responding by changes in their morphological characteristics at a concentration of this regulator in the medium of 0.01 g/ml or higher. The original collection strains in whose populations the variants were selected produced substances of the A-factor group. The A-factor-dependent variants differed in the level of the regulator required for maximal expression of the morphological characteristics: it was necessary to introduce the A-factor at a concentration of 1 g/ml for S. citreofluorescens and S. viridovulgaris subsp. albomarinus and at 10 g/ml for S. griseus.     

DNA homology and adsorption specificity of Pseudomonas aeruginosa virulent bacteriophages

Summary The DNA homology and adsorption specificity of newly isolated virulent bacteriophages of P. aeruginosa have been studied. On the basis of this analysis all phages were divided into four groups: k, m, mnP78-like and mnF82-like bacteriophages. DNA's of k as well as m phages were shown to possess different restriction patterns although they have an extensive homology. Unlike other groups, k phages were characterized by the presence of T4 DNA ligase-repaired, single-chain breaks.Abbreviations kbp kilobase pairs - EM electron microscopy     

Die bursa- und schwanzstrukturen und ihre aberrationen bei den strongylina (Nematoda) morphologische studien zum problem der pluri-und paripotenzerscheinungen

Zusammenfassung In der Einleitung ist das Ziel der Arbeit in den wesentlichsten Punkten herausgestellt.Die Bursastrukturen (Bursavelum und Rippen bzw. Papillen) der parasitischen Strongylina lassen sich von den entsprechenden Bildungen der freilebenden Rhabditina, vor allem der Gattung Rhabditis, ableiten und in ihren Einzelgliedern homologisieren.Die im Laufe der Phylogenie bei den Strongylina auftretenden strukturellen Transformationen lassen sich auf einige wenige, relativ einfache morphogenetische Grundvorgänge zurückführen, die da sind: Wachstumsallometrien, Materialkompensationen, Organverschmelzungen und Spaltungen (Fissationen), Rudimentationen und ähnliche Vorgänge.Innerhalb der Strongylina Bursa ist ein Gefälle der Wachstumsgradienten feststellbar, das sich vom Zentrum der Bursa sowohl nach distal als auch proximalwärts abschwdcht. Zunehmende Förderung der zentral gelegenen Organe (Rippen) führt zu entsprechender Reduktion der peripheren Bursastrukturen, was vor allem im terminalen Schwanzabschnitt auffällt und zur Ausbildung des oft nur noch als Rudiment vorhandenen Dorsalrippenkomplexes führt. Letzterer entspricht in seiner Gesamtheit der Schwanzspitze der peloderen Rhabditiden mit den Papillen 9 und 10.Die bei Rhabditis moist getrennten Papillen 7 und 8 sind bei allen Strongylina zu einer Rippe (Externodorsal-Rippe) verschmolzen, die jedoch in manchen Aberrationen durch Abspaltung eines akzessorischen Astes ihre wahre Natur (als Verschmelzungsprodukt) zu erkennen gibt (Atavismus).Da dieselben Transformationsvorgänge innerhalb der Strongylina mehrfach unabhängig voneinander wirksam geworden sind, treten bestimmte Strukturformen als Parallelbildungen in verschiedenen phylogenetischen Union auf (polytope Entstehung).Zahlreich untersuchte Bildungsabweichungen (Aberrationen), deren Bedeutung für die Morphologie kurz umrissen wird, erschöpfen sich in den gleichen strukturellen Transformationstypen, die auch bei der Evolution der verschiedenen Union der Strongylina nachweisbar sind. Die Aberrationen führen daher häufig zu Atavismen oder zu Parallelvariationen (homologe Variationen").Die Zahl der Umwandlungsmbglichkeiten (Potenzen) der Bursastrukturen innerhalb der Strongylina ist beschränkt (Paripotenz im Sinne Haeckers). Bestimmte Arten (und Entwicklungshnien) haben jeweils nur bestimmte Potenzen realisiert. Andere können jedoch latent (virtuell) im Kryptotypus vorhanden sein, ohne normalerweise in Erscheinung. zu treten. In bestimmten Aberrationen können sie jedoch plötzlich realisiert werden, so ihr latentes Vorhandensein demonstrierend (Pluripotenz).Wie lange bestimmte Potenzen in einer Gruppe erhalten bleiben konnen, verdeutlichen auch die Schwanzhocker weiblicher Nematoden, als zum Bauplan der Nematoden gehbrende Bildungen. Die Potenz zur Ausbildung dieser Strukturen kommt offensichtlich sehr vielen Nematoden-Arten zu, wird jedoch nur in relativ wenigen Fällen, aber innerhalb der verschiedenen Gruppen bald hier, bald dort (disjunkte Verbreitung), realisiert. Es handelt sich bei den Schwanzhöckern um rudimentäre Organe, die bei keiner Nematoden-Art mehr voll ausgebildet erhalten sind. Ihre Rudimentation beruht zum Teil auf Materialentzug, als Folge von Unkonstruktionen der Schwanzregion, wobei die Adultstadien zuerst betroffen werden (Aphanisie nach Sewertzoff).Bei den in Chiropteren parasitierenden Strongylacanthinae haben sich Schwanzhöcker noch bei allen Arten erhalten, was ein offensichtlich archaisches Merkmal darstellt. Bei anderen Nematoden, denen sie nur im Larvalstadium zukommen, treten sie wohl durch Fötalisation in seltenen Fällen auch bei den adulten Stadien wieder auf.Alle speziellen Bursaformen der Strongylina lassen sich durch relativ wenige und einfache Transformationsvorgänge aus einem durch Abstraktion gewonnenen diagrammatischen Typus ableiten (Prinzip der variablen Proportionen" nach Troll).Die typisierten Umwandlungsvorgänge decken sich weitgehend mit den von Remane allgemein gefaßten strukturellen Typen der Realmutationen. Da sie bei den beobachteten Aberrationen, deren Entstehung auf dem Wege über Realmutationen sehr wahrscheinlich ist, in homologer Weise auftreten, kann das innerhalb der Strongylina zu beobachtende Evolutionsphänomen auf Realmutationen zurückgeführt warden.Obwohl sich die untersuchten strukturellen Transformationen in dem systematisch relativ wait gefaßten Rahmen einer Unterordnung abspielen (transspezifische Evolution nach Rensch), handelt es sich bei der von uns bevorzugten Terminologie (nach Woltereck und Remane), unter Berücksichtigung des Charakters der Umwandlungen, doch nur um Vorgänge, die in den Bereich der Mikroevolution fallen.     

Plant regeneration from cytoplasmic hybrids of rice (Oryza sativa L.)

Summary We obtained cybrid plants by electrofusing -irradiated protoplasts of a cytoplasmic male-sterile line A-58 CMS (Oryza sativa L.) and iodoacetamide (IOA)-treated protoplasts of the fertile (normal) rice cultivar Fujiminori. The cybridity of the plants was confirmed by mitochondrial (mt) DNA restriction endonuclease, and plasmid-like DNA analyses, and by isozyme, cytological and morphological investigations. The chromosome number of the cybrid plants is 24.     

Genomic organization of human complement protein C8α and further examination of its linkage to C8β

Human C8 is one of five complement components (C5b, C6, C7, C8, C9) that interact to form the cytolytic C5b-9 complex on target membranes. It is composed of three nonidentical subunits (C8, C8, C8) encoded by separate genes. C8 and C8 are linked on chromosome 1p32, whereas C8 is located on 9q22.3-q32. In this study, overlapping genomic clones were isolated and used to decipher the organization of the human C8 gene. The gene contains at least 11 exons spanning 70kb of DNA. When compared to C6, C8 and C9, there is a remarkable similarity in genomic organization, consistent with amino acid sequence comparisons that suggest these proteins are ancestrally related. Regions of each protein that are structurally similar are encoded in exons of correspondingly similar lengths with highly conserved boundaries and phases. Availability of genomic sequence also facilitated a more detailed analysis of C8 and C8 linkage. Based on analysis of genomic digests with cDNA probes, the loci were previously reported to be physically linked (< 2.5kb) and in a 5- 3 orientation. In the present study, results obtained using exon-specific probes indicate the loci are not as closely linked as initially believed. Furthermore, they suggest that cDNA probes used earlier yielded misleading information because they encode exons that are distributed across large segments of genomic DNA.     

Microbial transformation of ginsenoside Rb1 by Rhizopus stolonifer and Curvularia lunata

Of 49 microbial strains screened for their capabilities to transform ginsenoside Rb₁, Rhizopus stolonifer and Curvularia lunata produced four key metabolites: 3-O-[-d-glucopyranosyl-(1,2)--d-glucopyranosyl]- 20-O-[-d-glucopyranosyl]-3,12, 20(S)-trihydroxydammar-24-ene (1), 3-O-[-d-glucopyranosyl-(1,2)--d- glucopyranosyl]-20-O-[-d-glucopyranosyl]-3,12, 20(S)-trihydroxydammar-24-ol (2), 3-O-[-d-gluco- pyranosyl-(1,2)--d-glucopyranosyl]-3, 12, 20(S)-trihydroxydammar-24-ene (3), and 3-O--d-glucopyranosyl-3, 12, 20(S)-trihydroxydammar-24-ene (4), identified by TOF-MS, ¹H- and ¹³C-NMR spectral data. Metabolites 1, 3 and 4 were from the incubation with R. stolonifer, and 1 and 2 from the incubation with C. lunata. Compound 2 was identified as a new compound.     

Comparison of the ability of β-glucosidases from almonds and Fusarium oxysporum to produce n-alkyl-β-glucosides in organic solvents

Summary The effect of water activity on the synthesis of n-alkyl-D-glucosides through condensation of glucose and n-alcohols has been studied using the commercially available almond -glucosidase and -glucosidase isolated from Fusarium oxysporum. The two enzymes exhibited a different water activity optimum. The specificity and alcohol reactivity of the two enzymes have also been investigated. Both enzymes prefer primary alcohols. -Glucosidase from F. oxysporum presents a higher affinity for primary alcohols with alkyl chain length of 4–6, whereas in the case of almond -glucosidase both initial velocity and yield decrease when the carbon chain length increases.     

Early development of the self-fertilizing mangrove killifish <Emphasis Type="Italic">Rivulus marmoratus</Emphasis> reared in the laboratory

The mangrove killifish Rivulus marmoratus was reared at 25°±1°C and 17ppt salinity from 0 to 100 days after hatching (DAH), and its early development was described by examining growth and morphometric parameters, meristic characters (vertebral and fin-ray counts), bone-cartilage development, and pigmentation. Growth was isometric for preanal length, head length, snout length, body depth, pectoral-fin length, dorsal-fin length, anal-fin length, and caudal-peduncle depth. Negative allometric growth was observed in eye diameter and gape size. Meristic counts (mean±SD) for vertebrae (34.2±0.4) and dorsal- (8.6±0.5), anal- (11.4±0.5), and caudal-fin rays (30.2±0.8) were complete at 0 DAH (n=5), whereas pectoral-fin rays and pelvic-fin rays were complete by 30 DAH (14.5±0.4, n=5) and 60 DAH (4.2±0.8, n=5). Full ossification of meristic elements proceeded in the following sequence: vertebrae (by 30 DAH), caudal-, dorsal-, and anal-fin rays (by 60 DAH), pectoral-fin rays (between 60 DAH and 100 DAH), and pelvic-fin rays (by 100 DAH). Both morphological characters and meristic counts indicate that this species can be considered to be a juvenile after 9.8mm in standard length (20 DAH).     

Action of rat liver Galβ1-4GlcNAc α(2-6)-sialyltransferase on Manβ1-4GlcNAcβ-OMe,GalNAcβ1-4GlcNAcβ-OMe,Glcβ1-4GlcNAcβ-OMe and GlcNAcβ1-4GlcNAcβ-OMe as synthetic substrates

Incubation of synthetic Man\1-4GlcNAc-OMe, GalNAc1-4GlcNAc-OMe, Glc1-4GlcNAc-OMe, and GlcNAc1-4GlcNac-OMe with CMP-Neu5Ac and rat liver Gal1-4GlcNAc (2-6)-sialyltransferase resulted in the formation of Neu5Ac2-6Man1-4GlcNAc-OMe, Neu5Ac2-6GalNAc1-4GlcNAc-OMe, Neu5Ac2-6Glc1-4GlcNAc-OMe and Neu5Ac2-6GlcNAc1-4GlcNAc-OMe, respectively. Under conditions which led to quantitative conversion of Gal1-4GlcNAc-OEt into Neu5Ac2-6Gal1-4GlcNAc-OEt, the aforementioned products were obtained in yields of 4%, 48%, 16% and 8%, respectively. HPLC on Partisil 10 SAX was used to isolate the various sialyltrisaccharides, and identification was carried out using 1- and 2-dimensional 500-MHz¹H-NMR spectroscopy.Abbreviations 2D 2-dimensional - CMP cytidine 5-monophosphate - CMP-Neu5Ac cytidine 5-monophospho--N-acetylneuraminic acid - COSY correlation spectroscopy - DQF double quantum filtered - HOHAHA homonuclear Hartmann-Hahn - MLEV composite pulse devised by M. Levitt - Neu5Ac N-acetylneuraminic acid - Neu5Ac2en 2-deoxy-2,3-didehydro-N-acetylneuraminic acid     

Cloning theClostridium thermocellum thermostable laminarinase gene inEscherichia coli: The properties of the enzyme thus produced

Summary We have cloned a 1.9-kb-long fragment ofClostridium thermocellum DNA which encodes laminarinase (EC 3.2.1.39). The enzyme hydrolyzes the -1,3-glucoside bonds in -1,3-and in mixed -1,3-1,4-polyglucans. The enzyme's optimum pH value is around 8.5, temperature optimum –70°C. PAGE-determined mol. weight –32 kDa.Abbreviations used CMC carboxymethyl cellulose - pNPC p-nitrophenyl D cellobioside - pNPLac p-nitrophenyl- D-lactoside - pNPG p-nitrophenyl D glucopyranoside - pNPGal p-nitrophenyl- D galactopyranoside - pNPXyl p-nitrophenyl- - D xylopyranoside - Ap ampicillin - SDS-PAGE SDS polyacrylamide gel electrophoresis