Isozyme polymorphism ofβ-glucosidase inAspergillus nidulans

Electrophoretic analysis of the distribution of various electromorphs at different -glucosidase zones was carried out in natural populations ofA. nidulans, theA. nidulans group, and various species belonging to the genusAspergillus from diverse geographical areas of India. The data show the existence of three segregating zones for -glucosidase, designated -GluI, -GluII and -GluIII. All three zones are present in wild isolates ofA. nidulans, and only two, i.e., -GluI and -GluIII, in theA. nidulans group and -GluII and -GluIII in different species ofAspergillus exceptA. terreus, A. flavus, andA. brevipes, where only -GluIII is present. Overall nine electromorphs are observed at -GluI, three at -GluII, and six at -GluIII zones, respectively, It can be concluded that there may be three structural genes for -glucosidase coding the three polymorphic zones inA. nidulans.This research work was supported by the Council of Scientific and Industrial Research (CSIR), New Delhi.     

The problem of transition from the chemical to the biological evolution: Some possible solutions

On the basis of evidence that several low-molecular-weight substances as well as enzymes are compartmentalised within the socalled soluble phase of the cell, and other considerations, it is argued that DNA may not contain information for certain types of organisation found in living cells. It may be necessary for a cell to possess the non-DNA-controlled organisation for performance of its minimum functions; such organisation would then also serve as a template for its appearance in the daughter cell. The problem of transition from chemical to biological evolution (that is, the formation of the first cell) may be essentially the problem of emergence of such intracellular organisation for which information may not reside in DNA. Two possible mechanisms through which this may have happened are stated.     

Host genes involved in the replication of single-stranded DNA phage ?K

Summary Using various replication mutants of E. coli, the host genes that participate in the replication of some K12-specific single-stranded DNA phages have been determined. Functional products of dnaE,-F,-G and -Z genes are required for the multiplication of K, whereas dnaA,-B,-C(D), H,-I and -P are dispensable for viral replication. In contrast with polB, recA, B, C, or xth functions, host rep activity is essential for K. At the restrictive temperature, the yield of K was markedly reduced in the ligts7 mutant and partially decreased in a polA ^ts strain. The phage K is thus less dependent on the host cells than X174 and A which require additionally the dnaB,-C(D) and -H functions. Replication of phage St-1 depends on dnaG and -Z gene products, but not on dnaP function. Although not much affected in polA ^ts host, growth of St-1 was significantly diminished in dnaF or ligts7 mutants.     

Measurement of the protein backbone dihedral angle φ based on quantification of remote CSA/DD interference in inter-residue 13C′(i − 1)-13Cα(i) multiple-quantum coherences

A novel triple-resonance NMR method is presented for the measurement of the protein backbone dihedral angle based on differential multiple-quantum relaxation induced by relaxation interference between ¹H(i)-¹³C(i) dipolar and ¹³C(i–1) (carbonyl) chemical shift anisotropy mechanisms. The method employs a simultaneous transfer of ¹⁵N magnetization to the inter- and intra-residue ¹³C carbons as well as the directly attached carbonyl carbon ¹³C. Results obtained on ¹³C,¹⁵N-labeled ubiquitin demonstrate the potential of the method.     

The effect of antibiotics on the inhibition of callus induction and plant regeneration from cotyledons of sugarbeet (Beta vulgaris L.)

Callus induction and plantlet regeneration from cotyledonary expiants of sugarbeet was observed utilizing two media formulations, MS and a modified MS termed RVIM both supplemented with 1.0 g/ml BAP as the sole growth regulator. Callus induction was genotype dependent The USDA line 8787 produced the highest response for callus induction followed by Betaseed 4587 and the USDA line C600. This order was conserved on both media formulations. Shoot induction was consistently higher averaging 32% from the RVIM formulation over the 3 genotypes compared to 25% from MS. The antibiotics geneticin, gentamycin, hygromycin, kanamycin and phleomycin were screened with the modified RV system utilizing Betaseed 4587. Callus growth was inhibited by levels of 50 g/ml geneticin, 150 g/ml gentamycin, 10 g/ml hygromycin, 150 g/ml kanamycin and 20 g/ml phleomycin. The results indicate that the concentrations of antibiotics used to inhibit callus induction will be sufficient for use as selectable markers in transformation experiments with Beta vulgaris.Abbreviations B₅ basal medium (Gamborg et al, 1968) - BAP N⁶-Benzylaminopurine - IBA Indole-3-butanoic acid - MS basal medium (Murashige and Skoog 1968) - RVIM modified MS basal medium (Freytag et al, 1988) - MES (2[N-Morpholino] ethanesulfonic acid     

Polymorphism of the HLA DR1 haplotype in the Israeli population investigated at the serological,cellular, and genomic levels

In the present report, we used serological, cellular, and restriction fragment length polymorphism (RFLP) to investigate the DR1 haplotype in the Israeli population. We describe an Israeli homozygous typing cell (HTC), HLA-DwLVA, which defines a new lymphocyte-activating determinant associated with Bw65, DR1 and distinct from Dwl. The parents of this donor, non-Ashkenazi Algerian Jews, are first cousins and share HLA-Cw8, Bw65, BfS, DR1, DQw1, DPw4. No specificity could be assigned to HLA-DwLVA using the 91 Ninth Workshop HTCs. Two families and forty unrelated DR1 individuals were studied with DwLVA and a panel of DR1/Dw1 HTCs. HLA-DwLVA showed segregation as a single determinant within families. This new specificity was present in 24 out of 40 (60%) unrelated DR1 individuals, indicating that in the Israeli population DwLVA is the main lymphocyte-defined determinant associated with the serologically defined DRI specificity, in contrast to non-Jewish Caucasoids where DR1 is significantly associated with Dw1. The vast majority of DwLVA-positive carriers were also Bw65 carriers, indicating that Bw65, DR1, DwLVA may represent a typical allele combination in the Israeli population. The RFLP analysis established the correlation of certain RFLPs with Dw1 and DwLVA. In addition, we describe a cluster of RFLPs that may correspond to a new Dw subtype associated with DR1, for which no serological and cellular reagents have been described so far.     

Antitumor effect of recombinant tumor necrosis factor-α against murine sarcomas at visceral sites: tumor size influences the response to therapy

Summary We examined the antitumor efficacy of rTNF- administration on established tumor at two visceral sites, lungs and liver. Treatment of B6 mice harboring multiple (>100 foci of 0.5 mm diameter) 10-day pulmonary macrometastases from the MCA-106 sarcoma, with dosages of rTNF- (5–10 g, single dose i. v.) that caused hemorrhagic necrosis and regression of a 6 mm MCA-106 s. c. tumor, had no impact on the number (or size) of lung nodules. Similarly, rTNF- failed to show an antitumor effect in B6 mice with advanced day 8 or 10 multiple (>100 foci of 0.5 mm diameter) hepatic metastases at single i. v. doses up to 20 g, as measured by either enumeration of residual liver nodules or survival. B6 mice injected s. c. with MCA-106 sarcoma and treated with rTNF- as a single i. v. dose on day 0, 3, 5, or 7 experienced marked tumor regression only after the day 7 rTNF- injection, when the tumor had achieved a size of 5–6 mm in diameter. Since tumor size appeared important for rTNF- susceptibility in vivo, we next induced a single hepatic tumor of the MCA-106 sarcoma by the direct injection of cells into the left lobe of the liver and treated these mice at day 10 when the nodule had achieved a size of 5–6 mm in diameter. Increasing doses of rTNF- (up to 8 g) given as a single i. v. injection resulted in increasingly greater reductions in hepatic tumor as well as significant survival benefit of the treated mice. Sites of regressing hepatic tumor exhibited central necrosis accompanied by polymorphonuclear leukocytes and lymphocytes. Collectively, these results show that rTNF- administration can mediate a significant antitumor effect on visceral tumor and suggest that tumor size is an important factor in rTNF- susceptibility not only for tumors growing at s. c. sites but also for those established at visceral sites.Howard Hughes Medical Institute Research Scholar Abbreviations used: rTNF-, recombinant tumor necrosis factor-; B6 mice, C57BL/6 mice; MCA, 3-methylcholanthrene; HBSS, Hanks' balanced salt solution; LAK, lymphokine-activated killer; rIL-2, recombinant interleukin-2     

The mitochondrially encoded "phantom ATPase" of S. cerevisiae. II. A heterogeneous mitochondrial ATPase population in situ.

Summary The mitochondrial ATPase from a PHO 1 mutant (OLI 2, PHO 1, OLI 4 region on mit DNA of S. cerevisiae) was further examined. A new purification method using Lysolecithin instead of Triton allowed us to solubilize and separate a heterogeneous ATPase population from PHO 1-mitochondria: the major abnormal fraction had extremely low oligomycin-sensitivity (but normal specific immunological reactivity), while a minor normal fraction (representing about 20% of the initial mitochondrial ATPase activity) had high sensitivity and affinity for oligomycin.Moderate urea treatment of PHO 1-mitochondria leads to partial loss of ATPase activity and a concomitant increase of oligomycin-sensitivity, suggesting that a heterogeneous ATPase population exists in situ in the mitochondrial membrane: part of the major abnormal ATPase fraction is selectively inactivated by urea, producing a concomitant enrichment in the initially minor normal ATPase fraction.If the minor normal ATPase fraction is the only one capable of in vivo ATP synthesis, the deficient but oligomycin-sensitive cell growth and oxidative phosphorylation in vitro are readily explained.Further structural studies are under way to ascertain whether the minor normal ATPase fraction is strictly identical to the wild type, in which case PHO 1 is a regulatory gene, or not, in which case PHO 1 is a structural gene.     

Auswirkungen des ?Sauren Regens“ (Waldsterben) auf eine Kohlmeisen-(Parus major-) Population in den Hochlagen des Harzes

Zusammenfassung Waldschäden als Folge des Sauren Regens sind in den Hochlagen des Harzes sehr deutlich sichtbar und spürbar. Diese (Zer-)Störung des Waldökosystems trifft auch die sehr häufige und sonst wenig empfindliche Kohlmeise. In zwei Nistkasten-Untersuchungsflächen von zusammen 25 ha in 800–920 m Höhe hat 1969–1997 der Bestand um 50% abgenommen, die Gelegegröße um 8%, der Bruterfolg erfolgreicher Paare um 18%, der Bruterfolg bei Berücksichtigung auch der erfolglosen Paare um 30% und der Gesamtbruterfolg auf der Gesamtfläche um 50%. Die Zahl von Brüten auf leerem Nest ist höher als in anderen Regionen. 1971–1987 betrug das durchschnittlich Mindestalter der brütenden Weibchen gleichmäßig 2,2–2,4 Jahre, 1993–1997 nur noch 1,15 Jahre. Als Hauptursache für diese Entwicklung wird die Änderung des Bodenchemismus durch den Sauren Regen mit seinen Folgen für die Bäume (Waldsterben), die Fauna (Nahrungsverknappung vor allem im Winter durch Abnahme der Spinnen) und die Physiologie der Vögel (Kalkmangel) diskutiert.

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Effects of acid rain on a population of Great Tit(Parus major) in the higher regions of the Harz Mountains

Summary The damage to forests through acid rain is patently obvious in the higher regions of the Harz Mountains. This disturbance in the forest's ecological system has also affected the frequently found and otherwise robust Great Tit. In two study areas with nest boxes (together covering 25 hectars at a height of 800–920 m a.s.l.) the population has been halved in the period between 1969 and 1997. The clutch-size has declined by 8%, the breeding results of successful pairs by 18% and the nesting success when unsuccessful pairs are included by 30%. The total nesting success of the whole area has declined by 50%. The number of birds breeding on empty nests is higher than in other regions. Between 1971 and 1987, the average minimum age of nesting females was constant 2.2–2.4 years. Between 1993 and 1997, however, the average minimum age was only 1.15 years. The changes in the chemistry of the soil due to acid rain with its consequences for the trees (forest damage Waldsterben), the fauna (reduction in food abundancy, particulary in winter due to a decrease in the number of spiders) and the physiology of the birds (lack of calcium) are discussed as the main reasons for this development.

     

Relations between Computational and Experimental Engineering Approaches to Molecules from Molecular Fragments

Computational techniques have become important tools in the preliminary stages of the design of new molecules. The mutual arrangements of interacting molecular parts and the required accessibility of important reactive groups on the peripheral regions of possible new molecules can be tested by computational means before the expensive and often complex synthetic methods are used to the actual construction of these molecules. There are some common features involved in the computational representation of molecular fragments and the synthetic methodologies used in the process of incorporating actual molecular fragments in such engineered molecules. One trend that appears to link these two approaches is based on the following observation: the greater local autonomy is shown by the calculated electron density contribution of a given molecular fragment, the more likely that this fragment can be regarded as a suitable building block of the molecule, and it is also more likely that there are convenient synthetic methods for the delivery of this fragment to its desired target location in the new molecule to be synthesized. For a precise formulation of this statement, a new concept, the degree of molecular fragment autonomy is introduced, using the inherent properties of molecular electron densities.     

Differences in N-acetyllactosamine synthesis between β-1,4-galactosyltransferases I and V

Unlike classical -1,4-galactosyltransferase (-1,4-GalT I), -1,4-GalT V (formerly IV*) has little activity towards 1 mM N-acetylglucosamine [Sato et al. (1998) Proc Natl Acad Sci USA 95: 472-477]. The human -1,4-GalTs I and V were expressed individually in Sf-9 cells by transfection of the full coding sequences, and their N-acetyllactosamine synthetase activities were determined towards different N-acetylglucosamine concentrations. Kinetic studies using the cell homogenates as an enzyme source revealed that -1,4-GalTs I and V possess Km values of 0.6 mM and 33 mM towards N-acetylglucosamine, and of 48 µM and 41 µM towards UDPGal, respectively. No significant inhibition of N-acetyllactosamine synthesis with -lactalbumin was observed for -1,4-GalT V but the significant inhibition with -lactalbumin was observed for -1,4-GalT I.     

Untersuchungen über Intersexualität beiGammarus duebeni (Crustacea,Amphipoda)

Zusammenfassung 1. In der vorliegenden Arbeit wird eine Analyse der intersexuellen Geschlechtsausprägung beiGammarus duebeni gegeben, die sich auf die Untersuchung der primären und sekundären Geschlechtsmerkmale, des Wachstums, der Häutungsfrequenz und des sexualbiologischen Verhaltens stützt. Das Auftreten von Intersexualität wird zu hormonalen und genetischen Faktoren, welche die Geschlechtsrealisation steuern, in Beziehung gesetzt.2. Zum besseren Verständnis der Besonderheiten intersexueller Entwicklung werden zunächst Morphologie und Differenzierung des Genitalapparates normalgeschlechtlicher Tiere beschrieben. In beiden Geschlechtern werden postembryonal männliche wie weibliche abführende Geschlechtsorgane angelegt, und in den Gonaden entstehen wahrscheinlich primär Oocyten. Mit Beginn der äußeren sexuellen Differenzierung werden im männlichen Geschlecht die Anlagen der Ovidukte, im weiblichen Geschlecht die Analgen der Vasa deferentia zurückgebildet, während die Anlagen der Vesicula seminalis erhalten bleiben. Eine männliche Determinierung ist von der Entwicklung und hormonalen Aktivität der androgenen Drüse abhängig.3. Nach dem Grad der Ausprägung weiblicher beziehungsweise männlicher Sexualcharaktere werden fünf verschiedene Intersexualitätstypen unterschieden: stark weibliche Intersexe, die habituell und funktionell völlig normalen gleichen, jedoch Calceoli und (oder) ein oder zwei Penispapillen aufweisen; schwach weibliche Intersexe, die etwas stärker maskulinisiert sind, im hinteren Bereich des Ovars testikuläres Gewebe ausbilden und durch mehr oder weniger differenzierte Anlagen der Vesicula seminalis, Vasa deferentia sowie meist rudimentäre androgene Drüsen gekennzeichnet sind; mittlere Intersexe, deren Gonaden aus ovarialen und testikulären Bezirken in nicht festgelegter Verteilung bestehen und die weibliche wie männliche ableitende Geschlechtswege, rudimentäre oder partiell entwickelte androgene Drüsen und Oostegiten mit normalem, unvollständigem oder fehlendem Borstenbesatz ausbilden; schwach männliche Intersexe, die durch eine Ovarregion im vorderen Abschnitt des Hodens, durch das Vorhandensein paariger Oviduktanlagen und durch Oostegiten ohne Randborsten charakterisiert sind; stark männliche Intersexe, die einen typisch männlichen Geschlechtsapparat und Oostegiten ohne Randborsten in normaler oder reduzierter Zahl besitzen.4. Hinsichtlich der Wachstumsintensität gleichen stark weibliche Intersexe normalen und stark männliche sowie schwach männliche Intersexe normalen . Einen mehr oder weniger intermediären Wachstumsverlauf weisen die mittleren und schwach weiblichen Intersexe auf. Auch die Größen- und Wachstumsrelationen des Propodus der 1. und 2. Gnathopoden liegen bei schwach weiblichen, mittleren und schwach männlichen Intersexen in gradueller Abstufung zwischen den Werten, die für das weibliche und männliche Geschlecht gelten.5. Stark weibliche, stark männliche und meist auch schwach männliche Intersexe sind fertil und zeigen normales weibliches beziehungsweise männliches Sexualverhalten. Schwach weibliche und mittlere Intersexe sind steril. Sie können nicht oviponieren, da ihre Ovidukte blind geschlossen bleiben. Mittlere Intersexe sind jedoch in Ausnahmefällen als geschlechtstüchtig. Wie anhand der Präkopulationsbereitschaft feststellbar ist, können sich mittlere Intersexe in Anwesenheit normalgeschlechtlicher Partner als gegenüber und als gegenüber verhalten. Diese geschlechtliche Ambivalenz wird als eine Erscheinung von relativer Sexualität gedeutet.6. Bezüglich der Häutungsfrequenz nehmen schwach weibliche und mittlere Intersexe eine weniger ausgeprägte, schwach männliche Intersexe eine stärker ausgeprägte Mittelstellung zwischen und ein. Die Häutungsfrequenz der schwach weiblichen und mittleren Intersexe ist nicht streng festgelegt. Sie kann bei mittleren Intersexen nachweislich durch die Gegenwart eines präkopulierenden reguliert werden, wobei eine Annäherung an die Häutungsfrequenz der erfolgt.7. Intersexe treten in natürlichen Populationen mit einer Häufigkeit von höchstens 0,5% auf. In Laborzuchten kann bei Selektion bestimmter Stämme der Anteil der Intersexe beträchtlich (10% und mehr) vergrößert sein.8. Durch Transplantation der androgenen Drüse in weibliche Tiere wurde experimentell eine Geschlechtsumwandlung in männliche Richtung ausgelöst, wobei verschiedene intersexuelle Zwischenstufen durchlaufen werden. Mitunter wird nur eine partielle Maskulinisierung erzielt, die etwa bis zum Stadium der mittleren Intersexualität führt. Auf Grund dieser und anderer Befunde wird das Zustandekommen intersexueller Geschlechtsausprägung auf eine rudimentäre oder unvollkommene Entwicklung der androgenen Drüse zurückgeführt. Die Entstehung der Intersexualität wird durch eine unzureichende Produktion des androgenen Hormons und durch Selbstdifferenzierung des Ovars erklärt.9. Anhand von Karyotypanalysen wurde die Chromosomenzahl mit 52 (= 2n) bestimmt. Hinweise für das Vorhandensein von Heterochromosomen liegen nicht vor; diese und andere Ergebnisse deuten auf eine polyfaktorielle Geschlechtsbestimmung hin.10. Die anGammarus duebeni gewonnenen Befunde werden unter vergleichenden Aspekten zu den Intersexualitätserscheinungen anderer Amphipoden in Beziehung gesetzt. Die Probleme der hormonalen Regulation der Geschlechtsfunktion werden eingehend diskutiert.

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Investigations on intersexuality inGammarus duebeni (crustacea, amphipoda)

A detailed analysis of intersexuality inGammarus duebeni, concerning morphology of primary and secondary sexual characters, growth, molting frequency and sexual behavior is presented. Based upon five different types a classification of intersexuality is given reflecting the degree of femaleness and maleness with respect to genital structures and secondary sex characters. Additional information is presented concerning the intermediary character of growth rates, molting frequency and sexual behavior in certain intersexual types. In intersexes the androgenic gland mediating the hormonal control of primary and secondary male characters is mostly in a rudimentary condition or only partially developed. According to sex reversal experiments and to the conceptions ofCharniaux-Cotton, intersexuality is interpreted inGammarus duebeni to be caused by reduced production of the androgenic hormone and by self-differentiation of the ovaries. On the basis of chromosomal studies and other genetic results, a polygenic sex determination is suggested.

     

Identification of T-cell receptor V β deletion mutant mouse strain AU/ssJ (H-2 q ) which is resistant to collagen-induced arthritis

Our laboratory is involved in investigating the role of T-cell receptor (Tcr) in collagen-induced arthritis (CIA). During these studies we found AU/ssJ (H-2 ^q ) mice to be resistant to CIA like SWR (H-2 ^q ), as compared with other H-2 ^q strains with wild-type Tcr like DBA/1 and B 10. Q. Upon screening with monoclonal antibodies F23.1 and KJ23a, AU/ssJ was found to be F23.1 negative (V 8 Tcr negative) and KJ23a positive (V 17a Tcr positive). Southern blot analysis on liver DNA using specific Tcr-V probes confirmed the deletion of V 8 gene family and also showed that AU/ssJ mice have deletions of V 9, V 13, V 12, and V 11 genes of Tcr. Further, these mice show a restriction fragment length polymorphism pattern with V 10, V 6, and V 17 probes similar to SWR mice as compared with 1310 mice. Since SWR and AU/ssJ are from different backgrounds, these studies indicate that specific variable region chain genes of Tcr are crucial for susceptibility to CIA in mice. Furthermore, these studies identify an additional inbred strain which has also deleted 50% of its Tcr-V genes.     

The Correcting Role of Autonomous 3" →="" 5"="" exonucleases="" contained="" in="" mammalian="" multienzyme="" dna="" polymerase="" complexes"="

A study was made of the correcting role of autonomous 3" 5" exonucleases (AE) contained in multienzyme DNA polymerase complexes of rat hepatocytes or calf thymocytes. DNA was synthesized on phage X174 amber3 or M13mp2 primer–templates, and used to transfect Escherichia coli spheroplasts. Frequencies were estimated for direct and reverse mutations resulting from mistakes made in the course of in vitro DNA synthesis. The error rate of the hepatocyte complex was estimated at 3·10^–6 with equimolar dNTP, and increased tenfold when proteins accounting for 70% of the total 3" 5" exonuclease activity of the complex were removed. The fidelity of DNA synthesis was completely restored in the presence of exogenous AE ( subunit of E. coli DNA polymerase III). Nuclear (Pol _n) and cytosolic (Pol _c) forms of DNA polymerase were isolated from calf thymocytes. The former was shown to contain an AE (TREX2) absent from the latter. As compared with Pol _c, Pol _n had a 20-fold higher exo/pol ratio and allowed 4–5 times higher fidelity of DNA synthesis. The error rate of DNA polymerase complexes changed when dNTP were used in nonequimolar amounts.     

Binding of the galactose-specificPseudomonas aeruginosa lectin,PA-I,to glycosphingolipids and other glycoconjugates

The carbohydrate-binding specificity ofPseudomonas aeruginosa lectin I (PA-I) in iodinated or biotinylated form was studied. A large number of glycosphingolipids, as well as some glycoproteins and neoglycoproteins were used as ligands. Also, inhibition by free saccharides of PA-I binding to glycosphingolipids was tested. It was found that the lectin binds most strongly to terminal and nonsubstituted Gal3Gal- or Gal4Gal-structures.Abbreviations PA-I Pseudomonas aeruginosa lectin I - Cer ceramide - lactosylceramide Gal4GlcCer - iso globotriaosylcerami Gal3Gal4GlcCer - globotriaosylceramide Gal4Gal4GlcCer - globoside or globotetraosylceramide GalNAc3Gal4Gal4GlcCer - Forssman glycolipid GalNAc3GalNAc3Gal4Gal4GlcCer - P1 glycolipid Gal4Gal4GlcNAc3Gal4GlcCer - lactoneotetraosylceramide Gal4GlcNAc3Gal4GlcCer - B5 glycolipid Gal3Gal4GlcNAc3Gal4GlcCer - gangliotetraosylceramide Gal3GalNAc4Gal4GlcCer - GM1 Gal3GalNAc4(NeuAc3)Gal4GlcCer - RBC red blood cells - BSA bovine serum albumin - PBS phosphate-buffered saline - SDS sodium dodecyl sulfate - TLC thin-layer chromatography - HPLC high pressure liquid chromatography - MS mass spectrometry - FAB fast-atom bombardment - EI electron impact     

Comparative residual toxicities of pesticides to the predator Agistemus industani (Acari: Stigmaeidae) on citrus in Florida

Residual toxicities of registered and selected experimental pesticides used on citrus against Agistemus industani Gonzalez (Acari: Stigmaeidae) were compared. Pesticides considered highly toxic to A. industani were: abamectin 0.15 EC at 731ml/ha+FC 435-66 petroleum oil at 46.8l/ha, pyridaben 75WP at 469g/ha, ethion 4EC at 7.01l/ha+FC 435-66 petroleum oil at 46.8l/ha, propargite 6.55 EC at 3.51l/ha, chlorfenapyr 2SC at 1.46l/ha applied alone or in combination with FC 435-66 petroleum oil at 46.8l/ha, sulphur 80DF at 16.81kg/ha, dicofol 4EC at 7.01l/ha, fenbutatin oxide 50WP at 2.24kg/ha, benomyl 50WP at 2.24kg/ha, benomyl 50WP at 1.68kg/ha+ferbam 76 GF at 5.60kg/ha, ferbam 76GF at 11.21kg/ha, neem oil 90EC at 46.8l/ha, and copper hydroxide DF (40% metallic copper) at 4.48kg metallic copper/ha+FC 435-66 petroleum oil at 46.8l/ha. Pesticides that were moderately to slightly toxic included: copper sulphate 98% at 4.48kg metallic copper/ha+FC 435-66 petroleum oil at 46.8l/ha, fenbuconazole 2F at 280ml/ha+FC 435-66 petroleum oil at 46.8l/ha, FC 435-66 petroleum oil applied alone at 46.8l/ha or 23.4l/ha, and diflubenzuron 25WP at 1.40kg/ha. Pesticides that were non-toxic included: fenbuconazole 2F at 585ml/ha, malathion 57EC at 5.85l/ha, FC 435-66 petroleum oil at 46.8l/ha, carbaryl 80S at 3.36kg/ha, chlorpyrifos 4EC at 4.68l/ha, and formetanate 92SP at 1.12kg/ha. Understanding the toxic effects of field weathered pesticides against key predacious mite species is important for effective IPM. The results of this study provide a comparison of direct and indirect toxic effects of various pesticides to A. industani under field conditions.     

Variation of amino acid concentrations in the medium of HU β-IFN and HU IL-2 producing cell lines

There are different requirements for the nutrient medium of various mammalian cell lines. We have determined the behaviour of the amino acid concentrations in the medium of two growing cell lines used for producing human interleukin 2 and human interferon constitutively. The experiments are based on a fermentation process with a bubble free cell culture aeration system with porous moving membranes, which allows production of high cell densities without foaming. We found interesting alterations in which the actual amino acid ratios are able to trigger consumption and production of a particular component depending on the supply of other possible replacements. Such data indicate the complicated biochemical network of synthesis, conversion and transport phenomena. Finally, we demonstrated the influence of product synthesis upon the amino acid requirements using as an example transformed hu IL-2- and hu -IFN-producing mouse L-cells.Abbreviations CTLL Cytotoxic T Cell Line - DME Dulbeccos Modified Eagle - FCS Foetal Calf Serum - Hepes N(2-Hydroxyethyl)piperazine-N'-2-ethanesulfonic acid - hu -IFN human -Interferon - hu IL-2 human Interleukin 2 - THF Tetrahydrofuran     

Unique sequences for two HLA-DRB1 genes expressed on distinct DRw6 haplotypes

We have used restriction fragment length polymorphism (RFLP) analysis and DNA sequencing to characterize two distinct DRB1 alleles expressed on DRw52 and DQw7-associated haplotypes but not readily defined by conventional DR serology. These two haplotypes, designated HLA-D HAG and PEV, react variably with DRw13(w6), DRw14(w6), and the more broad DR 3+6 antisera. Analysis of RFLP revealed that HLA-D HAG and PEV are associated with different DRw52 variants, and that HAG is indistinguishable from DRw18(3) haplotypes. Sequencing of the HAG and PEV DRB1 genes showed each to represent novel alleles. Nevertheless, these sequences show similarities with the other alleles of the DR5, w6, and w8 family. HAG (DRB1*1303) appears to have arisen either from two recombinational events involving at least three DRB1 sequences (DRB1*1101, DRB1*0803, DRB1*0401) or from a single recombinational event together with multiple point mutational events. PEV appears to represent a DRB1*1301-1302/DRB1*1101 recombinant allele, with recombination having occured in the region of bases 175 – 198. The results of this study suggest that the DRw52 family haplotypes is derived from a relatively restricted number of ancestral sequences, with diversity among DRB1 alleles within this family arising through gene conversion or recombination events.     

Carcinogenic potential and genomic instability of beryllium sulphate in BALB/c‐3T3 cells

Occupational exposure to beryllium (Be) and Be compounds occurs in a wide range of industrial processes. A large number of workers are potentially exposed to this metal during manufacturing and processing, so there is a concern regarding the potential carcinogenic hazard of Be. Studies were performed to determine the carcinogenic potential of beryllium sulfate (BeSO₄) in cultured mammalian cells. BALB/c3T3 cells were treated with varying concentrations of BeSO₄ for 72 h and the transformation frequency was determined after 4 weeks of culturing. Concentrations from 50–200 g BeSO₄/ml, caused a concentrationdependent increase (9–41 fold) in transformation frequency. Nontransformed BALB/c3T3 cells and cells from transformed foci induced by BeSO₄ were injected into both axillary regions of nude mice. All ten Beinduced transformed cell lines injected into nude mice produced fibrosarcomas within 50 days after cell injection. No tumors were found in nude mice receiving nontransformed BALB/c3T3 cells 90 days postinjection. Gene amplification was investigated in Kras, cmyc, cfos, cjun, csis, erbB2 and p53 using differential PCR while random amplified polymorphic DNA fingerprinting was employed to detect genomic instability. Gene amplification was found in Kras and cjun, however no change in gene expression or protein level was observed in any of the genes by Western blotting. Five of the 10 transformed cell lines showed genetic instability using different random primers. In conclusion, these results indicate that BeSO₄ is capable of inducing morphological cell transformation in mammalian cells and that transformed cells induced by BeSO₄ are potentially tumorigenic. Also, cell transformation induced by BeSO₄ may be attributed, in part, to the gene amplification of Kras and cjun and some BeSO₄induced transformed cells possess neoplastic potential resulting from genomic instability.     

Studies on aneuploids of Petunia

Summary The seven possible primary trisomics of Petunia (2 n= 14) located in the progenies of triploid, hypertriploid and hypotriploid plants were distinguished from one another and from diploid on the basis of cytological and morphological criteria. They were provisionally named as Oval, Semi, Slender, Pseudonormal, Arrow, Narrow and Giant. In three of the trisomics, the extra chromosome was identified for the first time at pachytene stage. Postpachytene studies revealed no precise relationship between the length of extra chromosome and the frequency of multiple association.