Effects of pressure on germination of seeds of wheat (Triticum aestivum cv. Barqai) in saline and in non-saline media

The effects of an increase in the absolute environmental pressure (air, N₂, O₂ or hydrostatic), up to 1 MPa, on the germination of wheat seeds and the survival of wheat seedlings were studied. Seeds were exposed to saline and non-saline media, in Petri dishes, on a double layer of filter paper. They were then introduced for different time periods into a pressure chamber and pressurized by the addition of N₂ to the chamber in the range of ambient to 1 MPa. Subsequently the seeds were left to germinate under normal atmospheric conditions. Seed germination and subsequent growth decreased during the first 6 h and then regained the control levels. Nevertheless, application of similar pressures to seeds which had been submerged under water was highly inhibitory. Such effects of pressure seem to be the result of flooding with water of some crucial intercellular spaces and a consequent disturbance of O₂ supply to the germinating embryo. The additional flood-water comprised only 1–3% of the total water content of 24-h-old seedlings. Sensitivity of the submerged seeds and the germinating seedlings to pressure varied with age and developmental stage. Highest sensitivity to pressure was obtained with 12 to 72-h-old submerged seedlings. Removal of the excess water after the pressure treatment restored the germinability of the seeds.     

Differential proto-oncogene mRNA induction from rats treated with peroxisome proliferators

After experimental treatment of rats with clofibrate or ciprofibrate, two peroxisomes proliferators with hypolipidemic activity, RNAs were prepared from liver, kidney, heart and brain; hybridization was done with DNA probes for c-myc and c-Ha-ras oncogenes and for cyanide insensitive Acyl CoA oxidase, a peroxisomal protein. c-myc mRNA is highly abundant in liver and at a lower extent in kidney, especially after treatment with ciprofibrate; clofibrate also allows a c-myc mRNA increase, but at a lower extent. c-Ha-ras, which is already expressed in all tested tissues from control animals, is stimulated by clofibrate and ciprofibrate treatments. Comparatively these compounds stimulate the cyanide insensitive Acyl CoA oxidase expression as well as they increase the somatic index of liver and kidney. From these experiments we suggest that hepatocarcinogenesis triggered by some hypolipidemic agents could be mediated by proto-oncogene mRNA level increase.     

Assessing DNA for fish identifications from reference collections: the good,bad and ugly shed light on formalin fixation and sequencing approaches

Natural history collections are repositories of biodiversity and are potentially used by molecular ecologists for comparative taxonomic, phylogenetic, biogeographic and forensic purposes. Specimens in fish collections are preserved using a combination of methods with many fixed in formalin and then preserved in ethanol for long-term storage. Formalin fixation damages DNA, thereby limiting genetic analyses. In this study, the authors compared the DNA barcoding and identification success for frozen and formalin-fixed tissues obtained from specimens in the CSIRO Australian National Fish Collection. They studied 230 samples from fishes (consisting of >160 fish species). An optimized formalin-fixed, paraffin-embedded DNA extraction method resulted in usable DNA from degraded tissues. Four mini barcoding assays of the mitochondrial DNA (mtDNA) were characterized with Sanger and Illumina amplicon sequencing. In the good quality DNA (without exposure to formalin), up to 88% of the specimens were correctly matched at the species level using the cytochrome oxidase subunit 1 (COI) mini barcodes, whereas up to 58% of the specimens exposed to formalin for less than 8 weeks were correctly identified to species. In contrast, 16S primers provided higher amplification success with formalin-exposed tissues, although the COI gene was more successful for identification. Importantly, the authors found that DNA of a certain size and quality can be amplified and sequenced despite exposure to formalin, and Illumina sequencing provided them with greater power of resolution for taxa identification even when there was little DNA present. Overall, within parameter constraints, this study highlights the possibilities of recovering DNA barcodes for identification from formalin-fixed fish specimens, and the authors provide guidelines for when successful identification could be expected.     

Social Class,Social Mobility and Risk of Psychiatric Disorder - A Population-Based Longitudinal Study

Objectives

This study explored how adult social class and social mobility between parental and own adult social class is related to psychiatric disorder.

Material and Methods

In this prospective cohort study, over 1 million employed Swedes born in 1949-1959 were included. Information on parental class (1960) and own mid-life social class (1980 and 1990) was retrieved from the censuses and categorised as High Non-manual, Low Non-manual, High Manual, Low Manual and Self-employed. After identifying adult class, individuals were followed for psychiatric disorder by first admission of schizophrenia, alcoholism and drug dependency, affective psychosis and neurosis or personality disorder (N=24 659) from the Swedish Patient Register. We used Poisson regression analysis to estimate first admission rates of psychiatric disorder per 100 000 person-years and relative risks (RR) by adult social class (treated as a time-varying covariate). The RRs of psychiatric disorder among the Non-manual and Manual classes were also estimated by magnitude of social mobility.

Results

The rate of psychiatric disorder was significantly higher among individuals belonging to the Low manual class as compared with the High Non-manual class. Compared to High Non-manual class, the risk for psychiatric disorder ranged from 2.07 (Low Manual class) to 1.38 (Low Non-manual class). Parental class had a minor impact on these estimates. Among the Non-manual and Manual classes, downward mobility was associated with increased risk and upward mobility with decreased risk of psychiatric disorder. In addition, downward mobility was inversely associated with the magnitude of social mobility, independent of parental class.

Conclusions

Independently of parental social class, the risk of psychiatric disorder increases with increased downward social mobility and decreases with increased upward mobility.     

Preferential recognition of epitopes on peroxynitrite-modified alpha-2-macroglobulin by circulating autoantibodies in rheumatoid arthritis patients

Abstract

Autoimmune responses against post-translationally modified antigens are a hallmark of several autoimmune diseases. In this work, we have studied the changes in alpha-2-macroglobulin (α₂M) upon modification by peroxynitrite. Furthermore, we have evaluated the immunogenicity of modified α₂M in experimental rabbits and rheumatoid arthritis (RA) patients. Peroxynitrite-modified α₂M showed disturbed microenvironment and altered aromatic residues under UV and fluorescence studies. Aggregation, reduction in β-sheet content, production of nitrotyrosine and shift in amide I and II bands were observed in the modified α₂M by polyacrylamide gel electrophoresis besides CD and FTIR spectroscopic analysis. The exposure of hydrophobic clusters and changes in contact positions were observed in ANS and ThT binding assays. Immunological studies using ELISA showed peroxynitrite-modified α₂M as highly immunogenic producing high titre of specific antibodies in immunized rabbits. Cross-reactivity studies revealed the polyspecificity of the elicited antibodies. Direct binding ELISA and competitive inhibition studies confirmed the presence of circulating antibodies in the sera of RA patients having high specificity towards the peroxynitrite-modified α₂M as compared to the native α₂M. Sera from healthy (normal) human subjects showed lower binding with the native and modified protein. This study confirms that peroxynitrite induces structural modifications in α₂M and makes it immunogenic. The presence of neo-antigenic determinants on modified α₂M with enhanced binding for circulating autoantibodies in RA patients could offer new possibilities for diagnosis and etiopathology of the disease.

Communicated by Ramaswamy H. Sarma     

Microbial maceration and carbonate dissolution on cold‐temperate shelves

Nummulites beaumonti d'Archiac & Haime, 1853, originally described from El Basatin, Gebel Mokattam, Greater Cairo, is redefined from a nummulitic bank at the base of the Maadi Formation from its type locality as neotype. Schaub (1981) described the species from Libya and the type example of this species is still unknown. N. discorbinus Schlotheim (1820) is described as neotype from a nummulitic bank in the Mokattam Formation, which is located in the site of the Citadel tombs, near the Salah El Din Citadel, Gebel Mokattam, Egypt. According to Schaub (1981), the type example of this species is also unknown. For this reason the establishment of the neotypes of these two widespread nummulites species: N. discorbinus (Late Lutetian) and N. beaumonti (Bartonian) is important and tackled in this paper. N. qurnensis n. sp. ( = N. sp. cf. beaumonti in Boukhary et al. 2002) is introduced as a new species from the lower Bartonian part of the Qurn Formation of Helwan. Biometrical studies are carried out on the three species to present up to date information on these Nummulites, with particular attention to their biostratigraphic correlation with the Shallow Benthic Zones (SBZ) of Serra-Kiel et al. 1998. The authors would like to stress the importance of a careful designation of a neotype to avoid the introduction of complicated and unnecessary taxonomic problems.     

Iron and nickel complexes with heterocyclic ligands: Stability,synthesis, spectral characterization,antimicrobial activity,acute and subacute toxicity

The synthesis and characterization by elemental analysis, emission atomic spectroscopy, TG measurements, magnetic measurements, FTIR, ¹H NMR, UV–visible spectra and conductivity of a series of iron (II) and nickel (II) complexes with two heterocyclic ligands (L¹(SMX): sulfamethoxazole and L²(MIZ): metronidazole) used in pharmaceutical field and with a new ligand derived benzoxazole (L³(MPBO): 2-(5-methylpyridine-2-yl)benzoxazole), were reported. The formulae obtained for the complexes are: [M(L¹)₂ Cl₂]·nH₂O, [M(L²)₂Cl₂(H₂O)₂]·H₂O and [M(L³)₂(OH)₂]·nH₂O. Stability constants of these complexes have been determined by potentiometric methods in water–ethanol (90:10, v/v) mixture at a 0.2 mol L^?1 ionic strength (NaCl) and at 25.0 ± 0.1 °C. Sirko program was used to determine the protonation constants as well as the binding constants of three species [ML₂H₂]²⁺, [ML₂] and [ML]²⁺. The antimicrobial activity of the ligands and complexes was evaluated in vitro against different human bacteria and fungi using agar diffusion method.Iron sulfamethoxazole complex showed a remarkable inhibition of bacteria growth especially on Staphylococcus aureus and P. aeruginosa. The iron metronidazole complex is active against yeasts especially on Candida tropicalis strain. Nickel complexes presented different antibacterial and antifungal behavior's against bacteria and fungal.The acute toxicity study revealed that the iron complexes are not toxic at 2000 mg/kg dose orally administrated.LD₅₀ for nickel complexes was determined using graphical method.No significant differences in the body weights between the control and the treated groups of both rat sexes in subacute toxicity study using for iron complexes. Hematological and clinical blood chemistry analysis revealed no toxicity effects of the iron complexes. Pathologically, neither gross abnormalities nor histopathological changes were observed for these complexes.     

Isolation and characterisation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) degrading actinomycetes and purification of PHBV depolymerase from newly isolatedStreptoverticillium kashmirense AF1

Streptoverticillium kashmirense AF1 with the ability to degrade a natural polymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) was isolated from municipal sewage sludge by soil burial technique. The PHBV film was degraded by the action of extracellular enzymes secreted by the microorganisms. Degradation of PHBV was evident by the formation of clear zones of hydrolysis on the polymer containing mineral salt agar plates. The extent of PHBV degradation increased up to 30 days of incubation. Maximum production of PHBV depolymerase was observed both at pH 8 and pH 7, 45 °C, 1% substrate concentration and in the presence of lactose as an additional carbon source. Two types of extracellular PHBV depolymerases were purified fromS. kashmirense AF1 by gel permeation chromatography using Sephadex G-75. The molecular weights of the two proteins were found to be 35 and 45 kDa approximately, as determined by SDS-PAGE. The results of the Sturm test also showed more CO₂ production as a result of PHBV degradation, in the test as compared to control. The present findings indicated the degradation capabilities ofS. kashmirense AF1.