RAPD (arbitrary primer) PCR is more sensitive than multilocus enzyme electrophoresis for distinguishing related bacterial strains.

The RAPD (random amplified polymorphic DNA) fingerprinting method, which utilizes low stringency PCR amplification with single primers of arbitrary sequence to generate strain-specific arrays of anonymous DNA fragments, was calibrated relative to the widely used, protein-based multilocus enzyme electrophoretic (MLEE) typing method. RAPD fingerprinting was carried out on five isolates from each of 15 major groups of Escherichia coli strains that cause diarrheal disease worldwide (75 isolates in all). Each group consisted of isolates that were not distinguishable from one another by MLEE typing using 20 diagnostic enzyme markers. In our RAPD tests, three or more distinct subgroups in each MLEE group were distinguished with each of five primers, and 74 of the 75 isolates were distinguished when data obtained with five primers were combined. Thus, RAPD typing is far more sensitive than MLEE typing for discriminating among related strains of a species. Despite their different sensitivities, the same general relationships among strains were inferred from MLEE and RAPD data. Thus, our results recommend use of the RAPD method for studies of bacterial population genetic structure and evolution, as well as for epidemiology.     

Screening for point mutations in exon 10 of the low density lipoprotein receptor gene by analysis of single-strand conformation polymorphisms: detection of a nonsense mutation — FH469→Stop

DNA from 40 unrelated familial hypercholesterolemia (FH) heterozygotes were subjected to analyses of single-strand conformation polymorphisms (SSCPs) of exon 10 of the low density lipoprotein receptor (LDLR) gene. Four different SSCP patterns were observed. The underlying mutations were characterized by DNA sequencing. Three of the patterns represented the three genotypes of a recently described sense mutation in codon 450. A method based upon the polymerase chain reaction (PCR) was developed to analyze this mutation. The frequencies of the wild-type (G at nucleotide 1413) and mutant (A at nucleotide 1413) alleles were 0.56 and 0.44, respectively. The fourth pattern was found in only one FH heterozygote and was caused by heterozygosity at nucleotide 1469 (G/A). Nucleotide 1469 is the second base of codon 469_Trp(TGG). The GA mutation changes this codon into the amber stop codon, and is referred to as FH_469Stop. The mutant receptor consists of the amino terminal 468 amino acids. Because the truncated receptor has lost the membrane-spanning domain, it will not be anchored in the cell membrane. FH_469Stop destroys an AvaII restriction site, and this characteristic was used to develop a PCR method to establish its frequency in Norwegian FH subjects. Two out of 204 (1%) unrelated FH heterozygotes possessed the mutation.     

Distribution,causal agents,and infection dynamic of emerging ink disease of sweet chestnut in Southern Switzerland

Emerging diseases caused by both native and exotic pathogens represent a main threat to forest ecosystems worldwide. The two invasive soilborne pathogens Phytophthora cinnamomi and Phytophthora × cambivora are the causal agents of ink disease, which has been threatening Castanea sativa in Europe for several centuries and seems to be re-emerging in recent years. Here, we investigated the distribution, causal agents, and infection dynamics of ink disease in southern Switzerland. A total of 25 outbreaks were identified, 19 with only P. cinnamomi, 5 with only P. × cambivora, and 1 with both species. Dendrochronological analyses showed that the disease emerged in the last 20–30 years. Infected trees either died rapidly within 5–15 years post-infection or showed a prolonged state of general decline until death. Based on a generalized linear model, the local risk of occurrence of ink disease was increased by an S-SE aspect of the chestnut stand, the presence of a pure chestnut stand, management activities, the proximity of roads and buildings, and increasing annual mean temperature and precipitation. The genetic structure of the local P. cinnamomi population suggests independent introductions and local spread of the pathogen.     

Exclusion cage studies on the impact of predation on Helicoverpa armigera in cotton

The impact of predation on Helicoverpa armigera was studied in four field cage exclusion trials on cotton in Kenya. H. armigera egg cohorts were introduced inside predator‐free and open control cages, and the impact of local predator populations on the cohort was examined. Fourteen days after inoculation, exclusion cages had four times more larvae than controls, indicating a strong impact of predation. Ants and Anthocoridae were the predominant predator groups. Exclusion cages had more damaged fruiting plant parts (squares, flowers and bolls) than the control. In the absence of predators, natural mortality of H. armigera was greater as cotton matured, and is likely to be linked to the host plant condition.     

Phenylsuccinate reduces KCL-induced release of GABA evidence for the participation of the ketodicarboxylate carrier in the biosynthesis of transmitter-GABA

Summary The present study investigates the effects of phenylsuccinate (PS), an inhibitor of the mitochondrial ketodicarboxylate carrier (KCC), on release of-aminobutyric acid (GABA), glutamate (Glu), glutamine (Gln), and glycine (Gly), induced by potassium chloride (KCl) and by cardiac arrest caused by a halothane overdose. Microdialysates were collected from the hippocampus of anaesthetized rats, and analyzed by HPLC. Continuous perfusion of 50 mM PS through the dialysis probe, reduces release of GABA induced by KCl (50 mM for 10 min through the dialysis probe) by up to 72%. In addition, PS abolished KCl-induced release of Glu. Release of GABA during cardiac arrest was not reduced by PS, whereas PS reduced release of Glu in the early stage of cardiac arrest. PS furthermore increased the basal level of Gln, and reversed a decrease of Gln induced by cardiac arrest.It is proposed that the KCC is present in GABA'ergic neurons of the rat hippocampus, and that GABA, released by KCl, can be synthesized in a KCC dependent manner. It is also suggested that ischemia-induced release of GABA, to some extent, has a non-transmitter origin. The results furthermore indicate that uptake of Gln into GABA'ergic and Glu'ergic neurons is not regulated by simple demand mechanisms.Abbreviations PS phenylsuccinate - KCC ketodicarboxylate carrier - GABA -aminobutyric acid - Glu glutamate - Gln glutamine - Gly glycine - -KG -ketoglutarate - Mal malate - KRB-buffer Krebs-Ringer bicarbonate-buffer - HPLC high pressure liquid chromatography     

Inter-twin relationships and mental health.

We evaluated dominance-submissiveness between co-twins and its relationship to mental health in a cohort study of 419 twins followed from pregnancy to 22-30 years of age. Dominance-submissiveness between co-twins was assessed from three separate perspectives: physical dominance, psychological dominance, and verbal dominance. Depressive, nervous, and psychosomatic symptoms were analyzed in different twin groups. In the physical domain, males were more commonly dominant than females at school age and in adulthood. Before and at school age, girls were more dominant than boys in the psychological and verbal domains, as well as in total dominance. These differences disappeared in adulthood, and 81% of adult twins felt themselves equal to their co-twin in total dominance. Submissiveness in the psychological domain seemed to be associated with increased depressiveness, nervous complaints and psychosomatic symptoms in males of male-female twin pairs. Verbally submissive males in same-sex twin pairs had more depression and psychosomatic symptoms. Among females of same-sex twin pairs, submissiveness in the psychological domain was most clearly associated with depressive symptoms, whereas psychological or verbal dominance-submissiveness among females from male-female twin pairs was not associated with symptoms. Psychologically dominant males and females of same-sex twin pairs expressed greater nervousness than did their co-twins. We conclude that being submissive, especially in the psychological domain, to a female twin partner seems to be stressful, whereas it is easier, especially for females, to be submissive to a male twin partner.     

A Sperm Midpiece Defect in a Hereford Bull with Variable Semen Quality and Freezability

A midpiece sperm defect with a frequency of 25-35% in ejaculates obtained from a Hereford bull with a 60 d non-retum rate of 76.4% after careful pre- and postfreeze semen selection was studied in light microscope and by transmission electron microscopy. The defect consisted in a folding and coiling of the distal midpiece characterized by disorganization and irregularity of mitochondria surrounding the axial fiber bundle, combined with retraction of doublet fibers and dislocation and fracturing of these elements and the corresponding dense fibers. Based on examination of the sper- matogenic epithelium it was concluded that the alterations in the axial fiber bundle were secondary to those in the mitochondrial sheath. The abnormality appeared to be related to the “Dag-like” defect earlier observed in different breeds.     

The Second Messenger, Cyclic AMP, Is Not Sufficient for Myelin Gene Induction in the Peripheral Nervous System

Abstract: The adenylyl cyclase-cyclic AMP (cAMP) second messenger pathway has been proposed to regulate myelin gene expression; however, a clear correlation between endogenous cAMP levels and myelin-specific mRNA levels has never been demonstrated during the induction or maintenance of differentiation by the myelinating Schwann cell. Endogenous cAMP levels decreased to 8–10% of normal nerve by 3 days after crush or permanent transection injury of adult rat sciatic nerve. Whereas levels remained low after transection injury, cAMP levels reached only 27% of the normal values by 35 days after crush injury. Because P₀ mRNA levels were 60% of normal levels by 14 days and 100% by 21 days after crush injury, cAMP increased only well after P₀ gene induction. cAMP, therefore, does not appear to trigger myelin gene induction but may be involved in myelin assembly or maintenance. Forskolin, an activator of adenylyl cyclase, increased endoneurial cAMP levels only in the normal nerve, and in the crushed nerve beginning at 16 days after injury, but at no time in the transected nerve. Only by treating transected nerve with 3-isobutyl-1-methylxanthine (IBMX), an inhibitor of cAMP phosphodiesterases, in combination with forskolin was it possible to increase cAMP levels. No induction of myelin genes, however, was observed with short- or long-term treatment with IBMX and forskolin in the transected nerve. A three-fold increase in phosphodiesterase activity was observed at 35 days after both injuries, and a nonmyelinated nerve was shown to have even higher activity. These experiments, therefore, suggest an important role for phosphodiesterase in the inactivation of this second messenger-dependent stimuli when Schwann cells are non-myelinating, such as after sciatic nerve injury or in the nonmyelinated nerve, which again implies that cAMP may be required for the maintenance of the myelin sheath.