Deletion mapping of a new gustatory mutant inDrosophila melanogaster

Summary A gustatory mutant ofDrosophila melanogaster insensitive to the taste of salt has been isolated. Genetic crosses and a deletion mapping analysis show that this mutation, designatedgust-M ₁ is located in the 93C_3–6-93D_6–7 region of the third chromosome.gust-M ₁ is also insensitive to the taste of quinine sulfate. The behavior of this mutant may be explained by assuming thatgust-M ₁ could be a mutation perturbing functions in the central nervous system affecting the responses to both compounds.     

Participation of the second extracellular loop of claudin-5 in paracellular tightening against ions, small and large molecules

Tight junctions control paracellular permeability. Here, we analyzed the impact of residues in the second extracellular loop (ECL2) of mouse claudin-5 on paracellular permeability. Stable expression of claudin-5_{wild type} in MDCK-II cells—but not that of mutants R145A, Y148A, Y158A or E159Q—increased transepithelial electrical resistance and decreased fluorescein permeation. Expression of claudin-5_Y148A, _Y158A or _E159Q enhanced permeability of FITC-dextran_10 kDa, which was unchanged in cells expressing claudin-5_{wild type} or claudin-5_R145A. In contrast, targeting to tight junctions, strand morphology and tight junction assembly were unchanged. It is concluded that R145 is unessential for trans-interaction of claudin-5, but necessary for tightening against small solutes and ions. The highly conserved residues Y148, Y158 and E159 in ECL2 of claudin-5 contribute to homo- and/or heterophilic trans-interaction between classic claudins and thereby tighten the paracellular space against ions, small and large molecules. These results provide novel insights into the molecular function of tight junctions.     

The possible involvement of protein kinase C and phospholipase A2 inHydra tentacle regeneration

The participation of protein kinase C (PKC) in the regeneration of tentacles ofHydra vulgaris was studied. Regeneration was induced by 1,2-sn-dioctanoyl-glycerol (diC₈) and the novel diterpenoidic diacylglycerol verrucosin B (VB), a potent PKC activator extracted from marine sources. VB substantially increasedHydra average tentacle number (ATN) at concentrations 10,000 times lower than those needed for diC₈ to exert an analogous effect. When both synthetic and natural VB analogues were tested, the structure/activity relationship found inHydra tentacle regeneration was identical to that known for DAG-induced activation of PKC in vitro. VB-induced increase of ATN was strongly counteracted by the PKC inhibitors sphingosine and A3, but was not synergic with a tenfold increase of extracellular Ca²⁺ concentration or with an increase of intracellular Ca²⁺ concentration obtained either with the ionophore A23187 or with thapsigargin. This suggested the involvement of a non-Ca²⁺-dependent PKC in VB-triggeredHydra tentacle regeneration. The involvement of phospholipase A₂ (PLA₂) activation inHydra regenerative processes was studied using the novel site-specific inhibitor of the enzyme, oleyloxyethylphosphorylcholine (OOPC), which brought about a striking inhibition of ATN in the low molar range. This effect was reversed by arachidonic acid (AA), while an enhancement of ATN was also observed with an inhibitor of AA uptake from membrane phospholipids, thus suggesting that PLA₂-catalysed liberation of AA is involved inHydra tentacle regeneration. OOPC also blocked verrucosin B-induced PKC-mediated enhancement of ATN, thus suggesting that this effect is also mediated by PLA₂ activation. ATN was increased also by compound 48/80, a direct activator of pertussis toxin-sensitive GTP-binding proteins, and this effect was counteracted by pertussis toxin pretreatment. None of the known AA cascade inhibitors exhibited an effect on ATN comparable to that exerted by OOPC, but, surprisingly, the cycloxygenase inhibitor indomethacin strongly enhanced ATN, thus suggesting that prostanoids might effect a negative control onHydra regenerative processes. This represents the first attempt so far reported to study the implication of more than one biochemical pathway as a signalling event in the hydroid regenerative processes.     

Production and characterization of antibodies cross-reactive with major aflatoxins

Antibodies cross-reactive with 4 major aflatoxins were demonstrated three weeks after immunization of rabbits with an immunogen which was prepared by conjugating aflatoxin B₃ to bovine serum albumin. Aflatoxin B₃ was first converted to its hemisuccinate before conjugation to the protein. Tritiated aflatoxin B₁ (AFB₁) was used as the marker ligand both for antibody titer determination as well as for analysis of antibody specificity. Competitive RIA revealed that the antibodies have good cross-reactivity with aflatoxins B₁, B₂, G₁, and G₂ when tritiated AFB₁ was used as the marker ligand. The concentrations causing 50% inhibition of binding of³H-AFB₁ to the antibodies by unlabeled aflatoxins B₁, B₂, G₁, G₂ and B₃ were found to be 0.25, 3.34, 0.32, 4.0 and 0.53 ng/assay, respectively. The antibodies could be used for simultaneous analysis of aflatoxins B₁ and G₁, two of the most important toxic metabolites produced byAspergillus flavus andA. parasiticus.     

Comparative aspects of structure and action of molluscan neuropeptides

A number of neuropeptides were isolated from the ganglia and muscles of molluscs, and their actions were examined. Diverse neuropeptides, in addition to several classical neurotransmitters, were suggested to be involved in the regulation of the anterior byssus retractor muscle ofMytilus. A wide structural variety of members of theMytilus inhibitory peptide family was observed in each of the generaMytilus, Achatina andHelix. Gly-Trp-NH₂, the C-terminal dipeptide fragment of the neuropeptide AGPWamide, showed a more potent action than the parent peptide in all of the muscles examined. Peptides related to some molluscan neuropeptides were found to be distributed interphyletically. Some neuropeptides containing ad-amino acid residue were found inAchatina andMytilus. These aspects of molluscan neuropeptides are thought not to be exceptional.     

Regulation of ammonia uptake inAspergillus nidulans

Summary The ammonia uptake inA. nidulans was found to be linear for about 20 min, and was proportional up to 1.5 mg/ml dry cell density. The transport of ammonia does not involve energy. Normal and biotin deficientA. nidulans showed an identical K_m-values of 10.26×10^–5 M ammonia for uptake. The uptake of ammonium ion has been shown to be regulated by the intracellular concentration of ammonia.Acknowledgment. The award of research fellowship by the Govt of India, Department of Atomic Energy, Bombay, to JDD is acknowledged.     

Aktivierung einer alkalischen Phosphatase

Summary -Glycerophosphatase prepared from the intestinal mucosa of the calf was purified by fractionated precipitation with alcohol. A further concentration of the enzyme activity was attained by electrophoresis.The activity of the purified enzyme solution was reduced to of its original value when dialysed for 48 hours atp _H 4.5. Atp _H 6 and atp _H 10.5 only a less pronounced decrease of the activity occurred.By addition of heat-inactivated-glycerophosphatase to the enzyme solution which was partly inactivated by dialysis atp _H 4.5 the activity of the latter was increased by about 100%.     

Age-dependent tolerance toBaculovirus in last larval instars of the codling moth,Cydia pomonella,L., induced either for pupation or for diapause

Summary LD₅₀ values as well as time-dependent parameters of granulosis virus infections were determined at different times during the last larval instar (L₅) of the codling moth,Cydia pomonella L., induced either for pupation or for diapause. A significant increase of tolerance to virus was found in 48-h-old L₅ induced for pupation, and 24 h later in L₅ induced for diapause.     

On the usefulness of macroeconomic forecasts as inputs to forecasting models

A forecasting model for y_t based on its relationship to exogenous variables (e.g. x?_t) must use x?_t, the forecast of x?_t. An example is given where commercially available x?_t's are sufficiently inaccurate that a univariate model for y_t appears preferable. For a variety of types of models inclusion of an exogenous variable x?_t is shown to worsen the y_t forecasts whenever x?_t must itself be forecast by x?_t and MSE (x?_t) > Var (x?_t). Tests with forecasts from a variety of sources indicate that, with a few notable exceptions, MSE (x?_t) > Var (x?_t) is common for macroeconomic forecasts more than a quarter or two ahead. Thus, either:

• (a) available medium range forecasts for many macroeconomic variables (e.g. the GNP growth rate) are not an improvement over the sample mean (so that such variables are not useful explanatory variables in forecasting models), and/or
• (b) the suboptimization involved in directly replacing x?_t by x?_t is a luxury that we cannot afford.

     

The X-ray analysis of the structure of rifamycin Y

Riassunto La struttura cristallina dellap-iodio anilide della rifamicina Y è stata determinata con metodi tridimensionali a temperatura ambiente. La cella elementare monoclina, gruppo spaziale P2₁, ha i parametria = 9,49,b = 18,96,c = 15,30 Å, = 90°48, e contiene due molecole.La costituzione e la configurazione della rifamicina Y, C₃₉H₄₇NO₁₅, risultano quindi determinate attraverso lo studio cristallografico della suap-iodio anilide. Anche molte informazioni sulla configurazione della rifamicina Y possono essere tratte dalla configurazione allo stato solido di questo suo derivato.

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A brief preliminary report on this structure was read at a meeting of the Accademia Nazionale dei Lincei, Rome, on 12th March 1966².     

Oxygen equilibrium ofPriapulus hemerythrin

Summary The hemerythrin-containing coelomic fluid ofPriapulus caudatus shows a relatively low O₂ affinity (half-saturation O₂ tension P₅₀=8 mm at 10 °C) and a low O₂ capacity (near 1 vol.%). O₂ affinity is independent of pH but shows a large temperature sensitivity. A major role as a continuous O₂ transporter seems to be excluded.Acknowledgments. A major part of this work was carried out at the Kristineberg Marine Laboratory, Fiskebäckskil (Sweden) and in the Zoophysiology Department, Aarhus Universitet (Denmark).     

Resistance to ethidium bromide inAspergillus nidulans

Summary A mutant ofAspergillus nidulans resistant to ethidium bromide was isolated and the semi-dominant gene responsible for this resistance was allocated on linkage group II at 17.42±3.05 units of recombination from thewA₃ gene. The gene also confers cross-resistance to acriflavin, malachite green and crystal violet. It was also shown that riboflavin is antagonistic to the toxic effect of ethidium bromide, at certain concentrations. The mechanisms which could be responsible for the toxic effect of this drug are discussed and compared with those of acriflavin. The use of theEtb ₁ gene in genetical analysis through the parasexual cycle is suggested.Acknowledgements. The authors are thankful to the National Council for the Development of Science and Technology (CNPq) for financial support through PIG-SIP 04/053 FAPESP.     

Avilamycin

Summary A strain ofStreptomyces viridochromogenes produced a new crystalline antibiotic, Avilamycin, related to but not identical with Curamycin and Exfoliatin. Avilamycin, C₆₃H₉₄O₃₅Cl₂, gave on solvolytic degradation the following products: dichloroisoeverninic acid, 2-deoxy-d-rhamnose, 2,6-di-O-methylmannose, 4-O-methylfucose,l-lyxose and 3,5-diacetoxy--caprolactone.     

Enrichment of ligands with molecular dockings and subsequent characterization for human alcohol dehydrogenase 3

Alcohol dehydrogenase 3 (ADH3) has been assigned a role in nitric oxide homeostasis due to its function as an S-nitrosoglutathione reductase. As altered S-nitrosoglutathione levels are often associated with disease, compounds that modulate ADH3 activity might be of therapeutic interest. We performed a virtual screening with molecular dockings of more than 40,000 compounds into the active site of human ADH3. A novel knowledge-based scoring method was used to rank compounds, and several compounds that were not known to interact with ADH3 were tested in vitro. Two of these showed substrate activity (9-decen-1-ol and dodecyltetraglycol), where calculated binding scoring energies correlated well with the logarithm of the k _cat/K _m values for the substrates. Two compounds showed inhibition capacity (deoxycholic acid and doxorubicin), and with these data three different lines for specific inhibitors for ADH3 are suggested: fatty acids, glutathione analogs, and cholic acids.     

Über das komplexchemische Verhalten von aromatischen Karbonsäurehydraziden

Summary A simple method to evaluate rough values of complex-formation constants has been developed. We have determinedk ₁*-values for Cu⁺² and aromatic hydrazides.k ₁* is suitable for comparison with biological results.     

Recombinant synthesis of mouse Zn3-β and Zn4-α metallothionein 1 domains and characterization of their cadmium(II) binding capacity

Genetic engineering, coupled with spectro scopic analyses, has enabled the metal binding proper ties of the α and β subunits of mouse metallothionein 1 (MT) to be characterized. A heterologous expression system in E.coli has led to high yields of their pure zinc-complexed forms. The cadmium(II) binding properties of recombinant Zn₄-αMT and Zn₃-βMT have been studied by electronic absorption and circular dichroism. The former binds Cd(II) identically to α fragments obtained from mammalian organs, showing that the recombinant polypeptide behaves like the na tive protein. Titration of Zn₃-βMT with CdCl₂ results in the formation of Cd₃-βMT. The addition of excess Cd(II) leads to Cd₄-βMT which, with the extra loading of Cd(II), unravels to give rise isodichroically to Cd₉-βMT. The effect of cadmium-displaced Zn(II) ions and excess Cd(II) above the full metal occupancy of three has been studied using Chelex-100. The Cd₃-βMT species is stable in the presence of this strong metal-chelating agent. Received 20 May 1997; received after revision 7 July 1997; accepted 9 July 1997     

Allele-specific PCR reveals thatCYP6D1 is on chromosome 1 in the house fly,Musca domestica

A cytochrome P450, termed P450lpr, is the major P450 responsible for pyrethroid resistance in the Learn-PyR (LPR) strain of house fly. Recently, the putative gene (CYP6D1) coding for P450_lpr has been sequenced from the LPR and aabys strains of house fly. Allele-specific polymerase chain reaction (ASPCR) was used for linkage group analysis with backcross progeny from the wild type LPR strain and a multiple marker strain (aabys). We found thatCYP6D1 is linked to chromosome 1. The possible role of regulatory or modifying genes responsible for elevated P450_lpr expression is discussed in relation to the chromosomal linkage ofCYP6D1.     

Interchange trisomics in pearl millet

Summary In the progeny of interchange heterozygotes ofPennisetum typhoides, 2 interchange trisomic plants were obtained. These were selfed over 2 generations and the cytology of the parents, s₁ and s₂ plants was studied. The types of multiple associations in the s₁ and s₂ generations differed from those of the parent. A significant decrease in mean chiasma frequency was noted in s₁ and s₂.Acknowledgment. The author is grateful to Prof. em. J. V. Pantulu, Department of Botany, Andhra University, Waltair, for his suggestions and encouragement.     

Ligand recognition by the I domain-containing integrins

Seven of the integrin α subunits described to date, α ₁ , α ₂ , α _L , α _X , α _d , α _M and α _E , contain a highly conserved I (or A) domain of approximately 200 amino acid residues inserted near the amino-terminus of the subunit. As the result of a variety of independent experimental approaches, a large body of data has recently accumulated that indicates that the I domains are independent, autonomously folding domains capable of directly binding ligands that play a necessary and important role in ligand binding by the intact integrins. Recent crystallographic studies have elucidated the structures of recombinant α _M and α _L I domains and also delineated a novel divalent cation-binding motif within the I domains (metal ion-dependent adhesion site, MIDAS) that appears to mediate the divalent cation binding of the I domains and the I domain-containing integrins to their ligands.     

Hypertension expérimentale et protéines sériques

Summary The authors investigated the distribution of serum proteins, especially the ₂ fraction, in rats with artificially induced hypertension (A), in similar rats made normotone by choline-free diet (B), and in a control group (C). A rise in the ₂ fraction of groupA andB was found, without a significant change of the other serum proteins.