Tick-borne thogoto virus infection in mice is inhibited by the orthomyxovirus resistance gene product Mx1.

We show that tick-transmitted Thogoto virus is sensitive to interferon-induced nuclear Mx1 protein, which is known for its specific antiviral action against orthomyxoviruses. Influenza virus-susceptible BALB/c mice (lacking a functional Mx1 gene) developed severe disease symptoms and died within days after intracerebral or intraperitoneal infection with a lethal challenge dose of Thogoto virus. In contrast, Mx1-positive congenic, influenza virus-resistant BALB.A2G-Mx1 mice remained healthy and survived. Likewise, A2G, congenic B6.A2G-Mx1 and CBA.T9-Mx1 mice (derived from influenza virus-resistant wild mice) as well as Mx1-transgenic 979 mice proved to be resistant. Peritoneal macrophages and interferon-treated embryo cells from resistant mice exhibited the same resistance phenotype in vitro. Moreover, stable lines of transfected mouse 3T3 cells that constitutively express Mx1 protein showed increased resistance to Thogoto virus infection. We conclude that an Mx1-sensitive step has been conserved during evolution of orthomyxoviruses and suggest that the Mx1 gene in rodents may serve to combat infections by influenza virus-like arboviruses.     

Compensatory substitutions and the evolution of the mitochondrial 12S rRNA gene in mammals

12S ribosomal RNA (rRNA) gene sequences from a suite of mammalian taxa (13 placentals, 4 marsupials, 1 monotreme), for which phylogenetic relationships are well established based on independent criteria, were employed to study the evolution of this gene. Phylogenetic analysis of 12S sequences produces a phylogeny that agrees with expectations. Base composition provides evidence for directional symmetrical substitution pressure in loops; in stems, base composition is much more even. Rates of nucleotide substitution are lower in stems than loops. Patterns of nucleotide substitution show an overall preference for transitions over transversions, with this difference more profound in stems than loops. Among different transversion pathways, there is a wide range of transformation frequencies. An analysis of compensatory substitutions shows that there is strong evidence for their occurrence and that a weighting factor of 0.61 should be applied in phylogenetic analyses to account for the dependence of mutations at stem positions relative to positions where changes are independent. Among stem variables (i.e., stem length, interaction distance, substitution rates, G+C content, and the percentage of bases that are paired), several significant correlations were discovered, but stem length and interaction distance are uncorrelated with other variables.      

Topology prediction for helical transmembrane proteins at 86% accuracy.

Previously, we introduced a neural network system predicting locations of transmembrane helices (HTMs) based on evolutionary profiles (PHDhtm, Rost B, Casadio R, Fariselli P, Sander C, 1995, Protein Sci 4:521-533). Here, we describe an improvement and an extension of that system. The improvement is achieved by a dynamic programming-like algorithm that optimizes helices compatible with the neural network output. The extension is the prediction of topology (orientation of first loop region with respect to membrane) by applying to the refined prediction the observation that positively charged residues are more abundant in extra-cytoplasmic regions. Furthermore, we introduce a method to reduce the number of false positives, i.e., proteins falsely predicted with membrane helices. The evaluation of prediction accuracy is based on a cross-validation and a double-blind test set (in total 131 proteins). The final method appears to be more accurate than other methods published: (1) For almost 89% (+/-3%) of the test proteins, all HTMs are predicted correctly. (2) For more than 86% (+/-3%) of the proteins, topology is predicted correctly. (3) We define reliability indices that correlate with prediction accuracy: for one half of the proteins, segment accuracy raises to 98%; and for two-thirds, accuracy of topology prediction is 95%. (4) The rate of proteins for which HTMs are predicted falsely is below 2% (+/-1%). Finally, the method is applied to 1,616 sequences of Haemophilus influenzae. We predict 19% of the genome sequences to contain one or more HTMs. This appears to be lower than what we predicted previously for the yeast VIII chromosome (about 25%).     

Phenotypic variability and therapeutic implications of Candida species in patients with oral lichen planus

We investigated the prevalence and phenotypic variation of Candida species in oral lichen planus (OLP) and the therapeutic implications of our findings. Eighty patients with clinically and histopathologically confirmed cases of OLP (64 non-erosive, 16 erosive) and a control group of 80 healthy individuals with no predisposing factors for oral candidiasis were examined for evidence of Candida infection. Oral swabs and smears were obtained for cytology and culture. Identification, speciation and antifungal susceptibility tests of Candida isolates were performed using an automated microbial identification system. Fifty percent of erosive OLP cases, 28% of non-erosive cases and none of the controls showed evidence of Candida. Candida albicans was found predominantly in non-erosive OLP, while other Candida species were predominate in erosive OLP. Non-Candida albicans isolates (C. glabrata, C. krusei) were resistant to the commonly used antifungals, clotrimazole and fluconazole. Candida infection is common in cases of OLP. We recommend antifungal sensitivity testing prior to antifungal therapy for the erosive form of OLP.     

Is there a relationship between branched amino acid pool size and cell cycle inhibition in roots treated with imidazolinone herbicides?

Excised pea root tips were cultured in White's medium for 24 h and then treated for 12 h with one of the imidazolinone herbicides at 0.2, 2, 20, or 200 M. Pursuit and Assert were almost ineffective in inhibiting the mitotic index (MI), except at the highest concentrations. Arsenal (ARS) and Scepter both showed good inhibition with 20 M by 8 h. Adding all three branched amino acids (BAA) (VAL, ILE, and LEU) at 0.1 mM blocked herbicide action. Treatment with the BAAs singly had no protective effect. Experiments were performed to determine the BAA pool size and MI after an 8-h treatment with ARS at 2 and 200 M and Chlorsulfuron (CS), a sulfonylurea herbicide, at 28 nM. Both CS and ARS at 200 M inhibited the MI to almost 0 by 8 h. ARS at 2 M inhibited the MI by about 40%. The BAA pool size in all three treatments was reduced by approximately 50%, whether the MI was totally blocked or not. The 1-mm root tips had a greater amount of VAL than did the mature portions of the roots, whereas ILE and LEU were slightly less in the root tip. Other soluble amino acids did not show consistent differences between herbicide-treated roots and controls. The implications of the pool size reduction, in instances where the MI was not totally inhibited, is discussed in light of new data from other laboratories on the mode of action of the imidazolinone herbicides.     

A comparison of two-point, three-point and deletion mapping in the C cistron of rhizobiophage 16-3, with an explanation for the recombination pattern

A general tendency for additivity prevailed in recombination frequencies for two-point fine-structure mapping of 14 mutants in the C cistron of Rhizobium meliloti phage 16–3, with little evidence of any marker effect. Intracistronic three-point mapping indicated that double crossovers are rare. Deletion mapping indicated that the two- and three-point mapping data gave the correct order of the mutations. A high frequency (5 to 8%) of c/c⁺ heterozygotic phage progenies was observed in standard crosses. This pattern implies formation of a relatively long region of heterozygosity. Together with the results of the three-point tests, it suggests certain properties of the branch migration and resolution steps envisioned in current mechanisms of recombination.     

Transferability of macrolide, lincomycin, and streptogramin resistances between group A, B, and D streptococci, Streptococcus pneumoniae, and Staphylococcus aureus

The transferability of plasmid pRI405 between various streptococci of groups A, B, and D, Streptococcus pneumoniae, and Staphylococcus aureus is described. pRI405 originated from Streptococcus faecalis and encodes for resistance to macrolides, lincomycin, and streptogramin B (MLS resistance). The host range of the well-documented streptococcal plasmid pAM beta 1 was found to be similar to that of pRI405. Cleavage with restriction enzymes suggests that pRI405 belongs to a related family of MLS resistance plasmids.     

Gemeinsame Aufzucht von fünf jungen Tannenmeisen (Parus ater) durch Trauerschn?pper (Ficedula hypoleuca) und Tannenmeisen

Summary A female Pied Flycatcher (Ficedula hypoleuca) built over a Coal Tits' (Parus ater) nest containing five chicks. Nevertheless, the nestlings survived on account of their parents' feedings, and because they were probably lifted up in the nest-box sitting on top of the material brought into the box by the flycatcher-. After nest-completion the Pied Flycatcher- layed four eggs, but refused breeding. Instead, and probably due to the intense begging of the five Coal Tit chicks, the Flycatcher-pair immediatly started to rear its young. Based on the good care the chicks were given by their parents and foster-parents they came along nicely and fledged some days later.     

Demonstration and topographical distribution of LHRH receptors in the central nervous system in the normal and castrated male rat

The topographical distribution of [125I]-LHRH binding sites was studied on brain sections of adult male rat by quantitative autoradiography. High density of sites was observed in the hippocampus, amygdala and entorhinal cortex (4-7 fmol of LHRH bound/mg protein). Lower density of sites was observed in the septum and frontal cortex. The receptor density was not significantly modified at day 5 following castration. Under the same conditions the pituitary receptors were significantly increased. The presence of specific LHRH binding sites in the limbic system may explain the behavioural effect observed following intracerebroventricular injection of LHRH. However, their functions under physiological conditions remain to be elucidated.