A genomic BAC library and a new BAC-GFP vector to study the holocentric pest Spodoptera frugiperda

Two genomic tools for the study of Lepidoptera and the holocentric structure of their chromosomes are presented in this paper. A bacterial artificial chromosome (BAC) library was constructed using nuclear DNA partially digested with HindIII from eggs of Spodoptera frugiperda. The library contains a total of 36,864 clones with an average insert size of 125 kb, which corresponds to approximately 11.5 genome equivalents. Hybridization screening of the library was performed with eight single-copy genes, giving an average hit of 10 clones per marker gene. Colinearity between the genome and BACs was demonstrated at the triose phosphate isomerase (tpi) locus. Probing of the library with a PCR fragment internal to the 18S ribosomal gene allowed an estimation of the rDNA locus size close to 115 repeats per haploid genome. A new vector (pBAC3.6eGFP) for transient transfection into S. frugiperda cell lines has been constructed. It is based on the BAC vector, pBAC3.6e, in which a gene encoding GFP was inserted under the control of the densovirus P9 promoter. This vector has the advantage to accommodate large genomic inserts and to be transfected in a large lepidopteran host range. It was used to construct a second BAC library from Sf9 cell nuclear DNA in order to allow a comparison between somatic and cell line genome organization.     

The characterization and molecular structure of hepatoproliferin: a liver regeneration factor from rat hepatocytes

Hepatoproliferin (HPF) was purified from regenerating rat livers as an oligomeric entity (big-HPF) from which the monomeric form (small-HPF) could be obtained using disaggregating conditions. By using a solid-phase ion-exchange method, small-HPF was forced to dissociate into two charged ionic species, namely norepinephrine (NE) and a sulfonated disaccharide with a molecular structure consisting of D-glucuronic acid bound to glucosamine 2,6-disulfate by a beta-glycosidic linkage having a beta, 1 --> 4 configuration. Monomeric HPF stemmed from the formation of three electrostatic bonds between the protonated amine groups of three norepinephrines, of which two bind to the deprotonated sulfonic groups of glucosamine 2,6-disulfate and one to the deprotonated carboxylic group of glucuronic acid, to constitute a tightly associated complex with a molecular mass of 1046 Da. This represents one of the two purified isoforms of small-HPF. The other isoform, which has a lower molecular mass of 877 Da, lack one NE, leaving the weaker carboxylic group of glucuronic acid unoccupied, to constitute a more acidic form of HPF.     

Detection of the pediocin gene <Emphasis Type="Italic">pedA</Emphasis> in strains from human faeces by real-time PCR and characterization of <Emphasis Type="Italic">Pediococcus acidilactici</Emphasis>UVA1

Background  

Bacteriocin-producing lactic acid bacteria are commonly used as natural protective cultures. Among them, strains of the genus Pediococcus are particularly interesting for their ability to produce pediocin, a broad spectrum antimicrobial peptide with a strong antagonistic activity against the food-borne pathogen Listeria monocytogenes. Furthermore, there is increasing interest in isolating new bacteriocin-producing strains of human intestinal origin that could be developed for probiotic effects and inhibition of pathogenic bacteria in the gut. In this work, we typed a new strain, co-isolated from baby faeces together with a Bifidobacterium thermophilum strain, and characterized its proteinaceous compound with strong antilisterial activity.     

The effect of pH on chemiluminescence of different probes exposed to superoxide and singlet oxygen generators

The compromised optima for high intensity chemiluminescence (CL), using superoxide generators, were all above pH 9.0 for the CL probes luminol and lucigenin. With luminol the optima were at pH 9.0 and 9.4 for the generators KO₂ and hypoxanthine/xanthine oxidase (HX/XO), respectively. Lucigenin, with the same generators, produced optima at pH 9.5 and 10.0, respectively. The probe methyl-Cypridina-luciferin analogue (MCLA) produced optima closer to neutral pH, which is preferred for physiological assessments. MCLA had optima at pH 6.0, 8.7 and 9.5 with KO₂ and with HX/XO optima at pH 4.8, 6.0, 7.0 and 8.7. When CL was assessed at physiological pH, MCLA observed superoxide radicals with a sensitivity of 100- and 330-fold more than luminol or luicigenin respectively. For singlet oxygen, the sensitivity of MCLA at this pH was 45- and 5465-fold more than for the said probes respectively. H₂O₂ did not elicit CL between pH 4 and 9.5 with any of the probes and did not influence the production of superoxide or singlet oxygen when co-assessed. Therefore CL could only be obtained when enzymes were used as converters. The optima for the enzyme-conversion system horseradish peroxidase (HRP)/H₂O₂, and luminol, were at pH 8.0 and 9.2. Lucigenin and HRP/H₂O₂ also had a biphasic CL profile with optima at pH 7.4 and 9.6. MCLA and HRP/H₂O₂ had five optima, with the major ones at pH 6.1 and beyond 10. The optima for the myeloperoxidase/H₂O system were at 8.6 and beyond 10.0 when luminol and 0.15 mol/L NaBr were used. © 1997 John Wiley & Sons, Ltd.     

SEX DETERMINATION IN DIPLOID-TRIPLOID CROSSES OF SPINACIA OLERACEA

Mahoney , D. L. Jules Janick , and E. C. Stevenson . (Purdue U., Lafayette, Ind.) Sex determination in diploid-triploid crosses of Spinacia oleracea. Amer. Jour. Bot. 46(5): 372–375. 1959.—Segregation for sex in spinach was studied in a series of diploid pistillate × triploid staminate crosses. The genetic data indicated that the functional gametes from staminate triploids were not confined to n and n + 1 types. Cytological analyses of progenies from a number of 2n ♀ × 3n (XYY and XYY) ♂ crosses revealed similar but non-homogeneous types of chromosome segregation. These crosses produced 49.3% diploids, 17.9% trisomies, 0.5% with 14 chromosomes, 1.2% with 16 chromosomes, 11.4% with 17 chromosomes, 18.9% triploids, and 0.8% with 19 chromosomes. The reciprocal crosses produced a higher percentage of aneuploid types including 1.7% with 15 chromosomes. Segregation for sex in each of the various chromosome numbered progeny of diploid-triploid crosses was presented and analyzed.     

Measurement of thoracoabdominal asynchrony: importance of sensor sensitivity to cross section deformations.

Discrepancies in the assessment of thoracoabdominal asynchrony are observed depending on the choice of respiratory movement sensors. We test the hypothesis that these discrepancies are due to a different dependence of the sensors on cross-sectional perimeter and area variations of the chest wall. First, we study the phase shift between perimeter and area (Phi(PA)) for an elliptical model, which is deformed by sinusoidal changes of its principal axes. We show that perimeter and area vary sinusoidally in the physiological range of deformations, and we discuss how Phi(PA) depends on the ellipticity of the cross section, on the ratio of transverse and dorsoventral movement amplitudes, and on their phase difference. Second, we compute the relationship between perimeter, area, and the output of the inductive sensor, and we proceed by comparing inductive plethysmography with strain gauges for several cross section deformations. We demonstrate that both sensors can provide different phase information for identical cross section deformations and, hence, can estimate thoracoabdominal asynchrony differently. Furthermore, the complex dependence of the inductive sensor on perimeter and area warns against this sensor for the evaluation of thoracoabdominal asynchrony.