Mating patterns of virgin and inseminatedDrosophila melanogaster of different alcohol dehydrogenase (Adh) genotypes

Summary Female choice mating experiments showed virgin femaleD. melanogaster of the 3Adh genotypes chose heterozygous (Adh ^F /Adh ^S ) males most commonly. Inseminated females chose mates randomly, but the likelihood of a female remating was genotypically dependent.Acknowledgment. Financial support was provided by the Australian Research Grants Commitee.     

Fungitoxicity ofm-fluorophenylalanine-containing peptides towardsPythium ultimum

Summary The tripeptide L-m-fluorophenylalanyl-L-alanyl-L-alanine was much more fungitoxic towardsPythium ultimum than the dipeptide L-m-fluorophenylalanyl-L-alanine orm-fluorophenylalanine. The fungitoxicity of the tripeptide was reduced by L-alanyl peptides and phenylalanine, but not by other amino acids. In contrast, the fungitoxicity ofm-fluorophenylalanine was unaffected by peptides, and was antagonized by several amino acids. These results suggest the effective delivery ofm-fluorophenylalanine into the cell by a tripeptide carrier.     

Development of DNA probes for cytotoxin and enterotoxin genes in enteric bacteria

Summary DNA probes to identify the genes encoding toxins in enteric bacteria have been developed. Use of these probes reduces the number of animals required for toxicity testing, as suspect bacteria can be directly tested for the presence of toxin. We have augmented the gene probes available by developing probes against theEscherichia coli enterotoxin LTII and shiga toxin fromShigella dysenteriae 1.The LTII gene fromE. coli 357900 was identified and characterised and a suitable internal probe was obtained. The LTII gene was found not to be common among enterobacteriae from various geographical locations. Isolates predominately of animal origin from Nigeria and Thailand hybridized with the probe.The shiga toxin gene was isolated fromS. dysenteriae 1 by a combination of in vivo and in vitro methods. An internal probe was identified and used against different serogroups ofShigella andE. coli isolated. The probe was found to hybridize withS. dysenteriae 1 isolates and also someS. flexneri andS. sonnei strains. Representatives were tested for toxin production and found to produce toxin at low levels.     

Identification of chicken lysozyme <Emphasis Type="Italic">g</Emphasis>2 and its expression in the intestine

Lysozyme is an important component of the innate immune system, protecting the gastrointestinal tract from infection. The aim of the present study was to determine if lysozyme is expressed in the chicken (Gallus gallus) intestine and to characterise the molecular forms expressed. Immunohistochemical staining localised lysozyme to epithelial cells of the villous epithelium along the length of the small intestine. There was no evidence for lysozyme expression in crypt epithelium and no evidence for Paneth cells. Immunoblots of chicken intestinal protein revealed three proteins: a 14-kDa band consistent with lysozyme c, and two additional bands of approximately 21 and 23 kDa, the latter consistent with lysozyme g. RT-PCR analyses confirmed that lysozyme c mRNA is expressed in 4-day, but not older chicken intestine and lysozyme g in 4- to 35-day chicken intestine. A novel chicken lysozyme g2 gene was identified by in silico analyses and mRNA for this lysozyme g2 was identified in the intestine from chickens of all ages. Chicken lysozyme g2 shows similarity with fish lysozyme g, including the absence of a signal peptide and cysteines involved in disulphide bond formation of the mammalian and bird lysozyme g proteins. Analyses using SecretomeP predict that chicken lysozyme g2 may be secreted by the non-classical secretory pathway. We conclude that lysozyme is expressed in the chicken small intestine by villous enterocytes. Lysozyme c, lysozyme g and g2 may fulfil complimentary roles in protecting the intestine.Received 4 August 2004; received after revision 1 September 2004; accepted 7 September 2004     

A new alarm pheromone (2-decen-1-yl-acetate) isolated from the stings ofApis dorsata andApis florea (Hymenoptera: Apidae)

Summary From sting extracts ofApis dorsata andA. florea, a substance was isolated which is active in alarm behaviour of both of these species but not ofA. mellifera andA. cerana. The active substance was identified as 2-decen-1-yl-acetate.     

Notable diversity in hemoglobin expression patterns among species of the deep-sea clam, <Emphasis Type="Italic">Calyptogena</Emphasis>

The deep-sea clams Calyptogena nautilei and C. tsubasa, which live in the cold-seep area at a depth of 3570 m in the Nankai Trough, Japan, have abundant hemoglobins (Hbs) in erythrocytes, similar to other Calyptogena species. We determined the cDNA-derived amino acid sequences of Hbs from two Calyptogena species. C. tsubasa was found to contain two dimeric Hbs, Hb I consisting of 145 amino acid residues and Hb II with 137 residues, similar to known Hbs from C. soyoae and C. kaikoi. Sequence identity was over 90% among the orthologous chains of Calyptogena Hbs. On the other hand, surprisingly, C. nautilei contained two monomeric Hbs, Hb III containing 141 residues and Hb IV with 134 residues. In addition, Hbs III and IV showed only 33–42% sequence identity with Hbs I and II from other Calyptogena species. The distal (E7) histidine, one of the functionally important residues of the heme protein, is replaced by glutamine in all Hb chains of Calyptogena species. A phylogenetic analysis indicated that C. nautilei Hb III is closer to Hb I from other Calyptogena species. We suppose that a Hb gene was duplicated at least three times in an immediate ancestor of Calyptogena and, presumably depending on physiological conditions different Hb sets are being expressed: dimeric Hbs I and II in C. soyoae, C. kaikoi and C. tsubasa, and monomeric Hbs III and IV in C. nautilei. Received 13 May 2003; received after revision 5 June 2003; accepted 12 June 2003     

Variance to mean ratio and the spatial distribution of animals

Summary Biological and statistical aspects of the application of variance to mean ratio to describe spatial distribution of animals are discussed. It is shown that the parameterb inTaylor's power laws ²=a m ^b shows intra-specific variation depending on the distribution of the constituent units of the population.a andb are only parameters of a very empirical way of describing the relation between variance and mean, which itself is an indicator for spatial distribution. Hence,a andb depend on the distribution behaviour of the animals, and not vice versa.Acknowledgments: The work was developed whilst I was at the Department of Entomology, University of Nairobi. I am grateful to ProfessorT. R. Odhiambo for his interest. I thank the referee for comments.     

Presenza di enteramina o di una sostanza enteraminosimile negli estratti gastrointestinali e splenici dei pesci e negli estratti gastroenterici delle Ascidie

Summary It was possible to demonstrate, by means of chemical and biological reactions, the presence of enteramine or of an enteramine-like substance in gastro-intestinal extracts of twoAscidi, Tethium plicatum andCiona intestinalis.The same substance was also found in gastric and still more in intestinal extracts ofSelacia, but seems not to be present in the spleen ofSelacia nor in the alimentary canal ofTeleostea.     

Control of obesity inA vy /a mice by 5α-androstan-17-one

Summary Treating VY/WfL-A ^vy /a mice with 5-androstan-17-one, a mammalian glucose-6-phosphate dehydrogenase inhibitor, prevented the mice from becoming obese. The weight difference between treated and controlA ^vy /a mice was mainly due to a decreased accumulation of triacylglycerol. The compound did not suppress appetite, had no detectable toxicity and did not affect the lipogenesis rates in the liver and carcass. The weight-controlling effect of 5-androstan-17-one inA ^vy /a mice was reversible upon withdrawal of treatment.The authors wish to thank Mr W.R. Gibson and Drs C.G. Culbertson and P.N. Harris for performing the pathological examinations.     

Beta-carotene affects gene expression in lungs of male and female <Emphasis Type="Italic">Bcmo1</Emphasis><Superscript>−/−</Superscript> mice in opposite directions

Molecular mechanisms triggered by high dietary beta-carotene (BC) intake in lung are largely unknown. We performed microarray gene expression analysis on lung tissue of BC supplemented beta-carotene 15,15′-monooxygenase 1 knockout (Bcmo1 ^−/−) mice, which are—like humans—able to accumulate BC. Our main observation was that the genes were regulated in an opposite direction in male and female Bcmo1 ^−/− mice by BC. The steroid biosynthetic pathway was overrepresented in BC-supplemented male Bcmo1 ^−/− mice. Testosterone levels were higher after BC supplementation only in Bcmo1 ^−/− mice, which had, unlike wild-type (Bcmo1 ^+/+) mice, large variations. We hypothesize that BC possibly affects hormone synthesis or metabolism. Since sex hormones influence lung cancer risk, these data might contribute to an explanation for the previously found increased lung cancer risk after BC supplementation (ATBC and CARET studies). Moreover, effects of BC may depend on the presence of frequent human BCMO1 polymorphisms, since these effects were not found in wild-type mice.     

Phytoalexin response is elicited by a pathogen (Rhizoctonia solani) but not by a mycorrhizal fungus (Glomus mosseae) in soybean roots

Summary A container system was constructed to study the response of soybean roots to infection by mycorrhizal or pathogenic fungi. The system allows a rapid and synchronous inoculation byGlomus mosseae orRhizoctonia solani. The phytoalexin glyceollin was measured in roots of inoculated and uninoculated plants for a period of 30 days. A significantly increased content of phytoalexin was found inR. solani-infected roots as compared to uninfected control roots. However, there was no difference in the glyceollin contents of the mycorrhizal and the control roots for up to 23 days after inoculation. The accumulation of glyceollin inR. solani-infected roots was not influenced by a subsequent inoculation withG. mosseae. Moreover glyceollin accumulated in mycorrhizal plants to the same extent as in control plants when they were inoculated withR. solani. The two fungi did not mutually influence the course of infection when they were inoculated together.     

<Emphasis Type="Italic">LAPTM4A</Emphasis> interacts with <Emphasis Type="Italic">hOCT2</Emphasis> and regulates its endocytotic recruitment

Human organic cation transporter 2 (hOCT2) is involved in the transport of endogenous and exogenous organic cations mainly in cells of the kidney and the brain. Here, we focus on the regulation of hOCT2 by direct protein–protein interaction. Screening within a mating-based split-ubiquitin-yeast-two-hybrid system (mBSUS) revealed the lysosomal-associated protein transmembrane 4 alpha (LAPTM4A) as a potential interacting protein. Interaction of LAPTM4A and hOCT2 was confirmed by pulldown assays, FRET microscopy analysis and immunofluorescence microscopy. Functionally, overexpression of LAPTM4A significantly decreased ASP⁺ uptake in HEK293 cells stably transfected with hOCT2, suggesting a negative regulation of hOCT2-mediated transport. Furthermore, overexpression of LAPTM4A leads to a significantly decreased hOCT2 plasma membrane expression in surface biotinylation experiments. In addition, significant expression of LAPTM4A in human kidney was demonstrated by immunoblotting and immunofluorescence.     

EGF-induced programmed cell death of human mammary carcinoma MDA-MB-468 cells is preceded by activation of AP-1

MDA-MB-468 is a human mammary adenocarcinoma cell line that overexpresses the epidermal growth factor (EGF) receptor and undergoes programmed cell death (apoptosis) in response to EGF treatment. Programmed cell death was shown to be greatly enhanced when cells were growth-arrested prior to EGF treatment. Apoptosis was characterized by an initial rounding up and detachment of the cells from their substrate starting about 12 h after EGF treatment, followed by chromatin condensation, nuclear fragmentation and oligonucleosomal fragmentation of the DNA at about 24 to 48 h. Cell death was dependent on de novo protein synthesis. We found a rapid induction of c-fos, c-jun and junB at the mRNA level after about 30 min of EGF treatment and a more delayed upregulation of fosB and fra-1. The junD gene was expressed in the absence of EGF, and it was moderately induced within 30 min of growth factor addition. The increase of the different fos and jun mRNAs were paralleled by an increase of activator protein-1 (AP-1) DNA binding activity. A characterization of the AP-1 complex revealed similar levels of several Fos and Jun proteins. Based on the kinetics of AP-1 accumulation and cell death, it seems likely that AP-1 contributes to the apoptotic cell death of EGF receptor-overexpressing MDA-MB-468 cells. Received 21 July 1997; received after revision 6 November 1997; accepted 6 November 1997     

Intergeneric interactions betweenEimeria separata (Apicomplexa) andNippostrongylus brasiliensis (Nematoda) in the rat

Summary Ova production inNippostrongylus brasiliensis infected rats was significantly greater than in rats singly infected with the helminth whenEimeria separata infections were introduced 4, 6 and 11 days postinoculation withN. brasiliensis. Patent periods were unaltered during concurrent infections. These results suggest that the presence ofE. separata affects helminth fecundity but does not increaseN. brasiliensis longevity as has been shown withE. nieschulzi.This study was supported by NIH MB RS Grant RRO8012-8.     

Mitochondrial DNA variation in social wasps (Hymenoptera,Vespidae)

Summary Patterns of restriction fragment length polymorphisms (RFLP) of European Vespinae were more similar within genera than between them. Distance trees were constructed that support the hypothesis of monophyly of the generaVespula andDolichovespula. Within the genusVespula, V. germanica was more closely related toV. rufa than toV. vulgaris. The position of the genusVespa remained uncertain due to the precision limits of the RFLP technique.     

Late DNA replication in the sex chromosomes ofDidelphis virginiana

Zusammenfassung Autoradiographisch wurde die späte DNS-Synthese derX- undY-Chromosomen vonDidelphis virginiana (Virginia Opossum) in Knochenmarkzellen untersucht. Die weiblichen Tiere zeigen typische späte Reduplikationsmuster in einemX, die männlichen das späte Reduplikationsmuster inX undY und inY allein.

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This study was supported in part by research grant No. AM-02504 and training grant No. Tl-AM-5277, National Institutes of Health. Dr.Reiss was a predoctoral trainee, National Institute of Arthritis and Metabolic Diseases, U.S. Public Health Service.     

Alexis Fontaine's ‘Fluxio-differential method’ and the origins of the calculus of several variables

Alexis Fontaine des Bertins (1704–1771) was the first French mathematician to introduce what we would now regard as results in the calculus of several variables. One example is Fontaine's theorem nF = (?F/?x)x + (?F/?y)y of 1737 for homogeneous expressions F of degree n in x and y. Many years later Fontaine indicated this particular result to have been ‘a continuation of the method of solution’ introduced by him in 1734 to solve the problem of the tautochrones. It is tempting to disregard this announcement, since the method applied to the tautochrones was a method of variations and not manifestly an exercise in the calculus of several variables. Do we have just another case of a mathematician's confusion about the origins of his earlier work? In this paper I describe Fontaine's possible intentions in his remarks.     

The role of bHLH genes in ear development and evolution: revisiting a 10-year-old hypothesis

In mouse ear development, two bHLH genes, Atoh1 and Neurog1, are essential for hair cell and sensory neuron differentiation. Evolution converted the original simple atonal-dependent neurosensory cell formation program of diploblasts into the derived developmental program of vertebrates that generates two neurosensory cell types, the sensory neuron and the sensory hair cell. This transformation was achieved through gene multiplication in ancestral triploblasts resulting in the expansion of the atonal bHLH gene family. Novel genes of the Neurogenin and NeuroD families are upregulated prior to the expression of Atoh1. Recent data suggest that NeuroD and Neurogenin were lost or their function in neuronal specification reduced in flies, thus changing our perception of the evolution of these genes. This sequence of expression changes was accompanied by modification of the E-box binding sites of these genes to regulate different downstream genes and to form inhibitory loops among each other, thus fine-tuning expression transitions.     

The occurrence of 5-hydroxytryptamine in the holothurian,Pentacter crassa

Summary 5-Hydroxytryptamine (5-HT) and hypoxanthine were isolated chromatographically from the holothurianPentacter crassa. This study was initiated as a result of the observed hypotensive activity of aP. crassa extract. This activity was also encountered in extracts of the holothuriansThelenota ananus andStichopus chloronatus and can be attributed to 5-HT.     

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%.