A method for the fractionation of the high-mobility-group non-histone chromosomal proteins

A method is given for the preparation of four non-histone chromosomal proteins, one of which, protein 14, hitherto has not been isolated. The method also enables the preparation of histone H1 in gram quantities. The four non-histone chromosomal proteins so prepared are all polar molecules over 50% of each being composed of acidic and basic amino acids. It is also shown that protein 14 can be prepared from calf thymus without prior isolation of chromatin.     

Characterisation of a haem active-site mutant of horseradish peroxidase, Phe41----Val, with altered reactivity towards hydrogen peroxide and reducing substrates.

A horseradish peroxidase variant ([F41V] HRP-C*), in which Val replaces the conserved Phe at position 41 adjacent to the distal His, has been constructed. Its composition and spectroscopic, catalytic and substrate-binding properties were compared with those of the wild-type recombinant (HRP-C*) and plant (HRP-C) enzymes. Presteady-state kinetic measurements of the rate constant for compound I formation (k1) revealed an eightfold decrease in the reactivity of the Phe41----Val variant towards H2O2, in comparison with HRP-C or HRP-C*. Measurement of the remaining rate constants, k2 and k3, for the two single-electron reduction reactions of [F41V] HRP-C with para-aminobenzoic acid as reducing substrate, showed that they were 2.5-fold and 1.3-fold faster, respectively. In contrast, analysis of data from steady-state assays with 2,2'-azinobis(3-ethylbenzthiazoline-6-sulphonate) as reducing substrate, showed decreased reactivity of the mutant enzyme to this compound, indicating a change in substrate specificity. Over the substrate range studied, the data for HRP-C* and for [F41V] HRP-C conformed to a simple modification of the accepted peroxidase mechanism in which a first-order step (ku), assumed to be product dissociation, becomes rate-limiting under our standard assay conditions. Calculations of rate constants from steady-state data yielded values of k1 for both enzyme forms in adequate agreement with those from pre-steady state measurements. They showed, furthermore, that both k3 for 2,2'-azinobis(3-ethylbenzthiazoline-6-sulphonate) and ku were substantially decreased, fivefold and tenfold, respectively, in the mutant. Analogous to the decrease in ku, we observed a twofold increase in the affinity of the mutant variant for the inhibitor benzhydroxamic acid. The coordination-state equilibrium of the haem iron also appeared shifted towards the hexacoordinate high-spin form. These observations indicate that in addition to affecting reactivity to H2O2, mutations in the distal region and close to the haem iron also affect reactivity towards different reducing substrates, inducing perturbations in the neighbourhood of the aromatic-substrate-binding site, known to be 0.8-1.2 nm from the haem iron.     

Serologic evidence of a Rickettsia akari-like infection among wild-caught rodents in Orange County and humans in Los Angeles County, California.

We detected antibodies reactive with Rickettsia akari, the etiologic agent of rickettsialpox in humans and in 83 of 359 (23%) rodents belonging to several species, collected in Orange County, CA. Reciprocal antibody titers >1:16 to R. akari were detected in native mice and rats (Peromyscus maniculatus, P. eremicus, and Neotoma fuscipes) and in Old World mice and rats (Mus musculus, Rattus rattus, and R. norvegicus), representing the first time that antibodies reactive with this agent have been detected in four of these species and the first report of these antibodies in rodents and humans west of the Mississippi River. We then tested serum samples from individuals who used a free clinic in downtown Los Angeles and found that 25 of 299 (8%) of these individuals had antibody titers >1:64 to R. akari. Serologic evidence suggested that R. akari or a closely related rickettsia is prevalent among several rodent species at these localities and that infection spills over into certain segments of the human population. Isolation or molecular confirmation of the agent is needed to conclusively state that R. akari is the etiologic agent infecting these rodents.     

Dream Imagery and Emotion.

The relationship between prominent visual imagery and emotion within dreams was investigated in relation to E. Hartmann's (1996) contextualizing image (CI) theory and M. Seligman and A. Yellen's (1987) dual imagery theory. Fifty-nine students recorded dreams over a 2-week period and submitted 115 dreams for analysis. Participants recorded ratings of emotion type and emotion intensity in each scene. Prominent visual images were identified and scored for intensity and detail by independent judges. As hypothesized from Hartmann's theory, there was a significant positive relationship between CI intensity and emotion intensity in the CI scene, emotion intensity generally peaked in the CI scene, and dreams containing a CI had higher overall ratings of emotion intensity than non-CI dreams. The result for the correlation of detail of prominent imagery with emotion was inconclusive, with a low positive correlation across CI scenes. This raises the possibility that the CI is not a unitary construct. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Tintinnids and other microzooplankton--seasonal distributions and relationships to resources and hydrography in a Maine estuary

Journal of Plankton Research, 9, 65–77, 1987. In Table II on page 70 the number of Tintinnopsis minuta foundat 0 m depth on 15 September 1981 should be 1200.     

Temperature-Light Intensity Effect on the Antagonism of Species of Cladosporium to Melampsora larici-populina on Cultivars of Populus X euramericana (Dode) Guinier

The antagonistic and colonistic effect of post-inoculation temperature and light intensity was investigated in a factorial experiment involving four Cladosporium spp., three poplar cultivars, two levels of temperature and light intensity, anda race of Melampsora larici-populina. The species of Cladosporium were more antagonistic to uredinial production on the inoculated leaf discs of the cultivars when these were incubated in high temperature – high light intensity than in any other regime. Whilst colonisation of the uredinia was usually minimal in discs incubated on the high temperature – high light intensity regime, the condition of incubation favouring maximum colonisation varied with the cultivar and the species of Cladosporium employed. All the major factors and most of their two-, three- and four-way interactions were significant determinants of variation in both antagonism to uredinial production and in colonisation of the uredinia produced. The validity ofextrapolation from such experiments to the field environment is discussed.     

Bacteriophage P22 as a vehicle for transducing cosmid gene banks between smooth strains of Salmonella typhimurium: use in identifying a role for aroD in attenuating virulent Salmonella strains

Summary A cosmid gene bank of the virulent Salmonella typhimurium C5 was constructed in Escherichia coli K12. The bank was repackaged into bacteriophage heads and transduced into the semi-rough S. typhimurium strain AS68 which expresses the LamB receptor protein. Approximately 6000 ampicillin-resistant transductants were pooled and used as host for the propagation of bacteriophage P22. The P22 lysate was able to transduce cosmid recombinants to smooth strains of S. typhimurium and individual transductants were selected which complemented various S. typhimurium auxotrophic mutations. A stable mutation was introduced into the aroD gene of S. typhimurium C5. The resulting aroD ^- mutant, named CU038, was highly attenuated compared with the wild-type parent strain and BALB/c mice immunised orally with CU038 were well protected against challenge with the virulent C5 parental strain. Using the cosmid bank repackaged into bacteriophage P22 heads it was possible to isolate cosmid recombinants that could complement the aroD mutation of CU038 either by in vitro selection using minimal medium or in vivo selection for restoration of virulence in BALB/c mice. Repackaged P22 cosmid banks could provide a simple system for selecting in vivo for Salmonella virulence determinants. A Salmonella typhi strain harbouring mutations in aroA and aroD was constructed for potential use as a live oral typhoid vaccine in humans.