"Candidatus Neoehrlichia chilensis" sp. nov.: Molecular detection and characterization of a novel Anaplasmataceae in wild rodents from Valdivia, southern Chile.

This study aimed to screen wild rodents from southern Chile, for the presence of Anaplasmatacea. Spleen samples from 33 wild rodents trapped in Valdivia Province were screened by conventional PCR (cPCR), targeting the Anaplasmataceae 16S rRNA gene (16S). Positive samples were further evaluated, targeting a larger 16S fragment, groEL operon, and gltA gene, followed by sequencing and phylogenetic analysis. Anaplasmataceae DNA was detected in 15% (five of 33) of the tested rodents (Abrothrix sp. [four of five] and Mus musculus [one of five]). Analysis of sequenced products based on the 16S gene revealed high similarity with "Ca. Neoehrlichia mikurensis," "Ca. Neoehrlichia lotoris" and "Ca. Neoehrlichia arcana" (97.8%-98.6%). A lower similarity was observed with Candidatus Neoehrlichia groEL (89.7%-92%) and gltA (79.5%-79.9%) loci. According to the 16SrRNA, groEL and gltA phylogenetic analyses, two closely related genotypes of "Candidatus Neoehrlichia" spp. from Chile were observed, which clustered together in a separate clade from other species in this genus. This study suggests the presence of two genotypes of a novel species of "Candidatus Neoehrlichia," proposed as "Candidatus Neoehrlichia chilensis," circulating in rodents from Chile. This is the first report of "Ca. Neoehrlichia" species in rodents from America.

Molecular epidemiology of hospital-onset methicillin-resistant Staphylococcus aureus infections in Southern Chile.

Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen of public health importance. In Chile, the Cordobes/Chilean clone was the predominant healthcare-associated MRSA (HA-MRSA) clone in 1998. Since then, the molecular epidemiological surveillance of MRSA has not been performed in Southern Chile. We aimed to investigate the molecular epidemiology of HA-MRSA infections in Southern Chile to identify the MRSA clones involved, and their evolutionary relationships with epidemic international MRSA lineages. A total of 303 single inpatient isolates of S. aureus were collected in the Valdivia County Hospital (2007-2008), revealing 33% (100 MRSA/303) prevalence for HA-MRSA infections. The SCCmec types I and IV were identified in 97% and 3% of HA-MRSA, respectively. All isolates lacked the pvl genes. A random sample (n = 29) of all MRSA was studied by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), SCCmec subtyping, agr and spa typing, and virulence genes profiling. PFGE analysis revealed the predominance (89%, 26/29) of pulsotype A and three additional pulsotypes, designated H1, I33, and G1. Pulsotype A (ST5-SCCmecI-spa-t149) is clonally related to the Cordobes/Chilean clone. Pulsotype H1 (ST5-SCCmecIVNT-spa-t002) is genetically related to the Pediatric clone (ST5-SCCmecIV). Pulsotype I33 (ST5-SCCmecIVc-spa-t002) is clonally related by PFGE to the community-associated MRSA (CA-MRSA) clone spread in Argentina, I-ST5-IVa-PVL(+). The G1 pulsotype (ST8-SCCmecIVc-spa-t024) is clonally related to the epidemic USA300 CA-MRSA. Here, we demonstrate the stability of the Cordobes/Chilean clone over time as the major HA-MRSA clone in Southern Chile. The identification of two CA-MRSA clones might suggest that these clones have entered into the healthcare setting from the community. These results emphasize the importance of the local surveillance of MRSA infections in the community and hospital settings.

IL-3 controls tau modifications and protects cortical neurons from neurodegeneration.

Interleukin-3 (IL-3) regulates the proliferation, survival and differentiation of haematopoietic cells via interaction with specific cell-surface receptors. IL-3 is expressed in several non-hematopoietic cell types. Studies have demonstrated the presence of IL-3 in the central nervous system, however, its physiological role in these cells is poorly understood. Previously we have been demonstrated that IL-3 prevents neuronal death induced by fibrillary ß amyloid in these cells, by PI 3-kinase and Jak/STAT pathway activation. In this study, we demonstrated that IL-3 significantly reduced Aß-promoted neurite degeneration and toxicity. Thus, this cytokine provides cellular protection against Aß neurotoxicity in primary cortical neuronal cells, by modulating microtubular dynamics and prevention of tau cleavage and hyperphosphorylation. We also demonstrates that IL-3 is expressed in the "in vivo" mouse model of AD, Tg2576, which also expresses human AßPP with the Swedish mutation. In summary, these results suggest that IL-3 could play a neuroprotective role in AD.

The protein kinase Cdk5. Structural aspects, roles in neurogenesis and involvement in Alzheimer's pathology.

A set of different protein kinases have been involved in tau phosphorylations, including glycogen synthase kinase 3beta (GSK3 beta), MARK kinase, MAP kinase, the cyclin-dependent kinase 5 (Cdk5) system and others. The latter system include the catalytic component Cdk5 and the regulatory proteins p35, p25 and p39. Cdk5 and its neuron-specific activator p35 are essential molecules for neuronal migration and for the laminar configuration of the cerebral cortex. Recent evidence that the Cdk5/p35 complex concentrates at the leading edge of axonal growth cones, together with the involvement of this system in the phosphorylation of neuronal microtubule-asociated proteins (MAPs), provide further support to the role of this protein kinase in regulating axonal extension in developing brain neurons. Although the aminoacid sequence of p35 has little similarity with those of normal cyclins, studies have shown that its activation domain may adopt a conformation of the cyclin-folded structure. The computed structure for Cdk5 is compatible with experimental data obtained from studies on the Cdk5/p35 complex, and has allowed predictions on the protein interacting domains. This enzyme exhibits a wide cell distribution, even though a regulated Cdk5 activity has been shown only in neuronal cells. Cdk5 has been characterized as a proline-directed Ser/Thr protein kinase, that contributes to phosphorylation of human tau on Ser202, Thr205, Ser235 and Ser404. Cdk5 is active in postmitiotic neurons, and it has been implicated in cytoskeleton assembly and its organization during axonal growth. In addition to tau and other MAPs, Cdk5 phosphorylates the high molecular weight neurofilament proteins at their C-terminal domain. Moreover, nestin, a protein that regulates cytoskeleton organization of neuronal and muscular cells during development of early embryos, and several other regulatory proteins appear to be substrates of Cdk5 and are phosphorylated by this kinase. Studies also suggest, that in addition to Cdk5 involvement in neuronal differentiation, its activity is induced during myogenesis, however, the mechanisms of how this activity is regulated during muscular differentiation has not yet been elucidated. Recent studies have shown that the beta-amyloid peptide (A beta) induces a deregulation of Cdk5 in cultured brain cells, and raises the question on the possible roles of this tau-phosphorylating protein kinase in the sequence of molecular events leading to neuronal death triggered by A beta. In this context, there are evidence that Cdk5 is involved in tau hyperphosphorylation promoted by A beta in its fibrillary form. Cdk5 inhibitors protect hippocampal neurons against both tau anomalous phosphorylations and neuronal death. The links between the studies on the Cdk5/p35 system in normal neurogenesis and its claimed participation in neurodegeneration, provide the framework to understand the regulatory relevance of this kinase system, and changes in its regulation that may be implicated in disturbances such as those occurring in Alzheimer disease.

[Autologous blood donation--good clinical practice and good recuperation practice]. / Eigenblutspenden--gute klinische Praxis und gute Herstellungspraxis.

Since October 1987, autologous blood donations have been performed in Bern and in 1994 reached a total of 946, representing 3% of all blood donations. This value is lower than the 4.9% for Switzerland as a whole. We report on the motivation and reasons of patients in favour of autologous blood donations and compare the results of 1989 with 1995. In the 6 years' period, the median patient's age increased from 54 to 63 years. The motivation for autologous blood donations changed from "routine surgical office" to "doctors". More than 50% of the patients mentioned the risk of acquiring an infectious disease, especially HIV, as the main reason for autologous blood donations. Labeling, testing or storage of autologous blood products of 37 out of 100 patients was incorrect, and in a another 20% an enquiry at the donation centres was needed to confirm the required quality of the autologous blood products. In 7 cases only one out of several blood bags was screened for viral diseases, and in 5 cases, unfortunately, none of the autologous blood products were tested for HIV or hepatitis B and C. Considering the negative cost-effectiveness of autologous blood transfusion, it is strongly recommended that the intrinsically low clinical benefit of autologous blood product should not be compromised by lack of good clinical practice and good manufacturing practice.