Yersinia spp. HMWP2, a cytosolic protein with a cryptic internal signal sequence which can promote alkaline phosphatase export.

The iron starvation-induced, 2,042-amino-acid protein HMWP2 of Yersinia enterocolitica has two internal hydrophobic segments which might promote its export and association with the cytoplasmic membrane. To determine whether part of HMWP2 could be exported beyond the periplasmic face of the cytoplasmic membrane, we used TnphoA mutagenesis to construct 10 hybrid proteins in which periplasmic alkaline phosphatase (PhoA) was fused to the end of C-terminally truncated HMWP1 (at amino acid positions 1751 and 1753 two independent isolates]) had high alkaline phosphate activity (close to that of the native enzyme), both in Escherichia coli and in Y. pseudotuberculosis, indicating that the PhoA segment of the hybrid reached the periplasm. Deletion studies showed that the export signal resides in the second hydrophobic segment of HMWP2. This result would be compatible with the topology of the protein in the cytoplasmic membrane predicted from the distribution of charged amino acids at either end of the two hydrophobic segments. However, two hybrids in which the junction was even further toward the C terminus of HMMWP2 (at positions 1793 and 1999) had only weak alkaline phosphatase activity, suggesting that the predicted topology is incorrect. The location of HMWP2 was therefore determined by subcellular fractionation. The results indicate that HMPW2 is mainly cytoplasmic, consistent with its presumed role in the ATP-dependent, nonribosomal synthesis of an unknown peptide. We propose that the high alkaline phosphatase activity associated with some of the HMWP-2-PhoA hybrids results from the unmasking of the cryptic export signal activity in the second hydrophobic segment of HMPW2.     

Understanding the Persistence of Plague Foci in Madagascar

Plague, a zoonosis caused by Yersinia pestis, is still found in Africa, Asia, and the Americas. Madagascar reports almost one third of the cases worldwide. Y. pestis can be encountered in three very different types of foci: urban, rural, and sylvatic. Flea vector and wild rodent host population dynamics are tightly correlated with modulation of climatic conditions, an association that could be crucial for both the maintenance of foci and human plague epidemics. The black rat Rattus rattus, the main host of Y. pestis in Madagascar, is found to exhibit high resistance to plague in endemic areas, opposing the concept of high mortality rates among rats exposed to the infection. Also, endemic fleas could play an essential role in maintenance of the foci. This review discusses recent advances in the understanding of the role of these factors as well as human behavior in the persistence of plague in Madagascar.     

Beiträge zur Entwicklungsgeschichte des sporogenen Gewebes der Gramineen und Cyperaceen. I.Zea mays

Zusammenfassung Auf Grund einer besonderen Zellteilngsfolge, die als mechanisch bedingt zu verstehen ist, ordnen sich die sporogenen Zellen beiZea mays so an, daß sie im Loculusquerschnitt wie die Sektoren eines Kreises um den Mittelpunkt herum liegen. Bei den folgenden Zellteilungen steht die Längsachse der Spindel den räumlichen Gegebenheiten entsprechend immer parallel, nie senkrecht, zur Antherenwand. Es erfolgen also keine Teilungen mehr in radialer Richtung, was im Zusammenwirken mit der Einstellung des Zellwachstums in antikliner Richtung bewirkt, daß die sporogenen Zellen im Zentrum des Antherenfaches auseinanderweichen und schließlich einen Hohlraum bilden.Die Pollenmutterzellen kleiden in einer Lage den Pollensack aus. Ihre Form ist ± stumpf-keilförmig, langgestreckt und die Längen ihrer drei Achsen verhalten sich ungefähr wie 321. Die längste Achse der Pollenmutterzellen und damit auch die Längsachse der Spindel der I. meiotischen Teilung steht immer parallel zur Antherenwand.Die Spindelanordnung der II. meiotischen Teilung erfolgt ebenfalls entsprechend den Raumverhältnissen. Die Spindeln stehen immer parallel zueinander, zur ersten Scheidewand und zur Antherenwand. Diese Art und Weise der Teilung führt dazu, daß die vier Zellen einer Tetrade sich in einer Ebene befinden und dem Tapetum anliegen. Auch nach der Isolierung der Mikrosporen wird diese Lage beibehalten.Die Auswirkung mechanischer Gesetzmäßigkeiten bei der Entwicklung des sporogenen Gewebes vonZea mays ist so stark, daß möglicherweise noch vorhandene andere Faktoren nur eine untergeordnete Rolle spielen.An dieser Stelle möchte ich Frau R.Wunderlich meinen besonderen Dank für mannigfache Beratung aussprechen.     

Ecological fitness, genomic islands and bacterial pathogenicity. A Darwinian view of the evolution of microbes

The compositions of bacterial genomes can be changed rapidly and dramatically through a variety of processes including horizontal gene transfer. This form of change is key to bacterial evolution, as it leads to ‘evolution in quantum leaps’. Horizontal gene transfer entails the incorporation of genetic elements transferred from another organism—perhaps in an earlier generation—directly into the genome, where they form ‘genomic islands’, i.e. blocks of DNA with signatures of mobile genetic elements. Genomic islands whose functions increase bacterial fitness, either directly or indirectly, have most likely been positively selected and can be termed ‘fitness islands’. Fitness islands can be divided into several subtypes: ‘ecological islands’ in environmental bacteria and ‘saprophytic islands’, ‘symbiosis islands’ or ‘pathogenicity islands’ (PAIs) in microorganisms that interact with living hosts. Here we discuss ways in which PAIs contribute to the pathogenic potency of bacteria, and the idea that genetic entities similar to genomic islands may also be present in the genomes of eukaryotes.     

The 102-Kilobase Unstable Region of Yersinia pestis Comprises a High-Pathogenicity Island Linked to a Pigmentation Segment Which Undergoes Internal Rearrangement

Several pathogenicity islands have recently been identified in different bacterial species, including a high-pathogenicity island (HPI) in Yersinia enterocolitica 1B. In Y. pestis, a 102-kb chromosomal fragment (pgm locus) that carries genes involved in iron acquisition and colony pigmentation can be deleted en bloc. In this study, characterization and mapping of the 102-kb region of Y. pestis 6/69 were performed to determine if this unstable region is a pathogenicity island. We found that the 102-kb region of Y. pestis is composed of two clearly distinct regions: an ≈35-kb iron acquisition segment, which is an HPI per se, linked to an ≈68-kb pigmentation segment. This linkage was preserved in all of the Y. pestis strains studied. However, several nonpigmented Y. pestis strains harboring an irp2 gene have been previously identified, suggesting that the pigmentation segment is independently mobile. Comparison of the physical map of the 102-kb region of these strains with that of strain 6/69 and complementation experiments were carried out to determine the genetic basis of this phenomenon. We demonstrate that several different mechanisms involving mutations and various-size deletions are responsible for the nonpigmented phenotype in the nine strains studied. However, no deletion corresponded exactly to the pigmentation segment. The 102-kb region of Y. pestis is an evolutionarily stable linkage of an HPI with a pigmentation segment in a region of the chromosome prone to rearrangement in vitro.     

Experimental-numerical analysis of minipig's multi-rooted teeth

The paper pertains to the analysis of the biomechanical behaviour of the periodontal ligament (PDL) by using a combined experimental and numerical approach. Experimental analysis provides information about a two-rooted pig premolar tooth in its socket with regard to morphological configuration and deformational response. The numerical analysis developed for the present investigation adopts a specific anisotropic hyperelastic formulation, accounting for tissue structural arrangement. The parameters to be adopted for the PDL constitutive model are evaluated with reference to data deducted from experimental in vitro tests on different specimens taken from literature. According to morphometric data relieved, solid models are provided as basis for the development of numerical models that adopt the constitutive formulation proposed. A reciprocal validation of experimental and numerical data allows for the evaluation of reliability of results obtained. The work is intended as preliminary investigation to study the correlation between mechanical status of PDL and induction to cellular activity in orthodontic treatments.