Borrelia afzelii gene expression in Ixodes ricinus (Acari: Ixodidae) ticks

Spirochetes belonging to the Borrelia burgdorferi sensu lato (s. l.) complex have evolved remarkable ability to survive in diverse ecological niches during transmission cycles between ticks and vertebrate hosts by variable gene expression. To understand the events during spirochete transmission from feeding ticks to hosts, mRNA levels of selected B. afzelii genes (bbk32, dbpA, ospA, ospC and vlsE) were measured by quantitative real-time SYBR Green PCR. B. afzelii infected Ixodes ricinus nymphs fed on laboratory BALB/c mice for 0, 24, 48, and 72 hours. The mRNA levels of the constantly expressed flagellin gene were used for the relative quantification of selected genes. Differences in gene expression profiles were observed in unfed ticks and during tick feeding. mRNA levels of bbk32 and dbpA showed distinctive decreasing patterns during the first 24 hours post-attachment, while ospC and vlsE mRNA levels increased significantly during the feeding process. In contrast, ospA levels decreased for the 48 hours of tick feeding and slightly increased by 72 hours. More detailed and comprehensive studies on regulation of gene expression in different Borellia genospecies on the vector-host interface would aid to develop effective strategies in preventing pathogen transmission.     

Risk factors associated with snoring in women with special emphasis on body mass index: a population-based study

STUDY OBJECTIVES: Habitual snoring may be regarded as an indicator of sleep-disordered breathing, and the health consequences of sleep-disordered breathing are well-known. The aim of this study was to analyze the risk factors associated with habitual snoring in a large sample of women, with special emphasis on the determinants of snoring in women with different body mass index (BMI) levels.Design and setting: A cross-sectional, epidemiologic, population-based study was performed by using a postal questionnaire that was sent to a randomly selected sample of 6,817 women >/= 20 years of age in Uppsala, Sweden. RESULTS: The total prevalence of self-reported habitual snoring was 7.6%. There was a clear age dependence, with the highest prevalence of habitual snoring (14%) occurring between the ages of 50 and 59 years. Self-reported habitual snoring was related to BMI, neck circumference, and smoking >/= 10 cigarettes a day, after adjusting for possible confounders. When analyzing the influence of different risk factors in separate BMI groups, the results varied among the groups. The influence of alcohol dependence on snoring frequency was only significant in women with a BMI of < 20 kg/m(2), while physical inactivity was only associated with habitual snoring in women with a BMI of >/= 30 kg/m(2). CONCLUSIONS: The prevalence of self-reported habitual snoring in women was strongly dependent on age and BMI. The importance of other risk factors differed depending on BMI, with alcohol dependence being associated with self-reported snoring in lean women, whereas physical inactivity was a risk factor for self-reported snoring in women with a high BMI.     

Exposed and concealed antigens as vaccine targets for controlling ticks and tick-borne diseases

Tick vaccines derived from Bm86, a midgut membrane-bound protein of the cattle tick, Boophilus microplus, are currently the only commercially available ectoparasite vaccines. Despite its introduction to the market in 1994, and the recognized need for alternatives to chemical pesticides, progress in developing effective antitick vaccines (and ectoparasite vaccines in general) is slow. The primary rate-limiting step is the identification of suitable antigenic targets for vaccine development. Two sources of candidate vaccine antigens have been identified: 'exposed' antigens that are secreted in tick saliva during attachment and feeding on a host and 'concealed' antigens that are normally hidden from the host. Recently, a third group of antigens has been distinguished that combines the properties of both exposed and concealed antigens. This latter group offers the prospect of a broad-spectrum vaccine effective against both adults and immature stages of a wide variety of tick species. It also shows transmission-blocking and protective activity against a tick-borne pathogen. With the proliferation of molecular techniques and their application to vaccine development, there are high hopes for new and effective antitick vaccines that also control tick-borne diseases.     

Genetic diversity patterns at the human clock gene period 2 are suggestive of population-specific positive selection

Period 2 (PER2) is a key component of the mammalian circadian clock machinery. In humans, genetic variation of clock genes or chronic disturbance of circadian rhythmicity has been implied in the onset of several phenotypes, ranging from periodic insomnias to advanced or delayed sleep phases, to more severe disorders. Peculiar geographic diversity patterns in circadian genes might represent an adaptive response to different light/dark cycles or environmental changes to which different human populations are exposed. To investigate the degree and nature of PER2 gene variation in human populations of different geographic origin, and its possible correlation with different latitudes, we sequenced a 7.7 kb portion of the gene in 20 individuals worldwide. In total, 25 variable sites were identified. The geographic distribution of haplotypes defined by five polymorphic sites was analyzed in 499 individuals from 11 populations from four continents. No evidence for latitude-driven selective effects on PER2 genetic variability was found. However, a high and significant difference in the geographic distribution of PER2 polymorphisms was observed between Africans and non-Africans, suggesting a history of geographically restricted natural selection at this locus. In support of this notion, we found several signals for selection in the sequences. The putative selected haplotype showed a recent coalescent age (8.7 Kyr), and an unusually high frequency in non-African populations. Overall, these findings indicate that a human clock-relevant gene, PER2, might have been influenced by positive selection, and offer preliminary insights into the evolution of this functional class of genes.     

Dobrava hantavirus causes hemorrhagic fever with renal syndrome in central Europe and is carried by two different Apodemus mice species

In central Europe, hemorrhagic fevers with renal syndrome (HFRS) in humans are caused by the hantavirus species Puumala (transmitted by voles) and a second, Hantaan-related species (transmitted by mice). The second virus could be identified as Dobrava virus. To date, 19 clinical cases of Dobrava infection have been found in Germany and Slovakia. All patients exhibited a mild/moderate clinical course and no case fatality occurred. Screening for infected rodents revealed that the striped field mouse (Apodemus agrarius) represents the main reservoir for Dobrava virus in central Europe. Nucleotide sequence comparisons and phylogenetic analysis based on complete and partial genomic S segment nucleotide sequences placed the Slovakian A. agrarius-derived hantavirus strains within the Dobrava species, forming a cluster on the Dobrava phylogenetic tree. In east Slovakia, a single Dobrava virus-infected yellow-necked mouse (Apodemus flavicollis) was trapped in a locality that predominantly showed Dobrava-infected A. agrarius. Comparison of the S segment sequence (nucleotides 381-935) revealed that the Dobrava strain from A. flavicollis shows only 84.3% nucleotide homology to A. agrarius-derived strains from this location but 96.3% homology to A. flavicollis-derived Dobrava strains from the Balkans (southeast Europe). Phylogenetic analysis of the partial S segment placed the A. flavicollis-derived Dobrava strain from Slovakia on a distinct Dobrava lineage (DOB-Af) together with the south-east European A. flavicollis-derived strains. The results indicate that Dobrava strains from A. agrarius (DOB-Aa) vs. A. flavicollis (DOB-Af) could develop different degrees of virulence in humans.     

Identification of the target of monoclonal antibody A6H as dipeptidyl peptidase IV/CD26 by LC MS\MS

The monoclonal antibody (MAb) A6H, originally developed to fetal renal tissues, was found to be highly reactive to renal cell carcinoma and was subsequently demonstrated to co-stimulate a subpopulation of T cells. The A6H antigen had not been identified heretofore. Antigen from detergent extracts of renal cell carcinoma cells (7860) was immunoabsorbed with A6H-agarose, and the resin-bound proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The antigen had a molecular weight of approximately 120 kDa as determined by Western blots. The 120-kDa protein band was excised and subjected to in-gel tryptic digestion, and the resulting peptides were separated and analyzed by liquid chromatography tandem mass spectrometry (LC MS\MS). The tandem mass spectra of the eluting peptides were used in combination with the SEQUEST computer program to search a human National Cancer Institute (NCI) protein database for the identity of the protein. The target antigen was shown to be dipeptidyl peptidase IV (DPP IV), which is also known as the cluster differentiation antigen CD26. Flow analysis of the expression of the A6H antigen and of CD26 on 7860 cells and on peripheral blood lymphocytes supported the identification of the A6H antigen as DPP IV. Recognition that the A6H antigen is DPP IV/CD26 afforded the opportunity to compare previous studies on A6H with those on other anti-CD26 antibodies in terms of expression in cancer cell lines and various tissues and as co-stimulators of T-cell activation.     

Association of the -159 CD14 gene polymorphism and lack of association of the -308 TNFA and Q551R IL-4RA polymorphisms with severe chronic periodontitis in Swedish Caucasians

BACKGROUND: Severe forms of periodontitis are suggested to have a genetic basis. OBJECTIVE: The aim of the present investigation was to study the association of gene polymorphisms related to some immune regulation components (G-308A TNFA, Q551R IL-4RA and C-159T CD14) with severe chronic periodontitis. MATERIALS AND METHODS: Sixty patients (aged 36-74 years; mean 54.5+/-8.5) with severe and generalized chronic periodontitis were included. The patients exhibited bone loss >50% at all teeth. Thirty-nine periodontally healthy subjects between 35 and 78 years of age (mean 51.0+/-10.9) were recruited as controls. DNA was isolated from peripheral blood cells and genotyping was performed by combination of PCR and restriction endonuclease mapping. RESULTS: While gene polymorphisms for TNFA and IL-4RA did not show any association with severe chronic periodontitis, the analysis of the -159 CD14 gene polymorphism revealed significant differences between test and control groups. The proportion of subjects that exhibited the TT genotype was significantly smaller in the group with severe periodontitis than in periodontal healthy group (p=0.028; Fisher's exact test). The C allele carriage was 90% in the periodontitis group and significantly higher than in the healthy control group (72%). CONCLUSION: It is suggested that the -159 CD14 gene polymorphism is associated with chronic periodontitis in Caucasian subjects of a north European origin.     

Immediate loading of implants placed with the osteotome technique: one-year prospective case series

Background: There is limited information regarding marginal bone‐level changes around immediately loaded implants placed with the osteotome technique. The aim of this case series is to prospectively evaluate the clinical and radiographic outcome of immediately loaded implants placed with the osteotome technique over a 12‐month period. Methods: Eighteen patients in need of oral prosthetic rehabilitation that included single implant placement in positions #4 to #13 and/or #20 to #29 participated in this prospective trial. A modified implant installation procedure with an under preparation of the implant bed using the osteotome technique and immediate loading of the implant was performed. Clinical examinations were performed at 2 weeks, 6 months, and 12 months of follow‐up. Radiographic examinations were performed at implant installation and at the 6‐ and 12‐month follow‐up visits. Results: One implant failed to integrate and was removed at 3 months after implant installation. Four of 20 implants had insertion torque value >35 Ncm. The mean marginal bone loss was ‐0.09 mm at the 6‐month and ‐0.19 mm at the 12‐month follow‐up visits. Conclusion: The present case series indicates that implants placed with the osteotome technique and immediately loaded did not demonstrate a high insertion torque and exhibited minimal marginal bone loss.     

When genetics and genealogies tell different stories-maternal lineages in Gaspesia

Data from uniparentally inherited genetic systems were used to trace evolution of human populations. Reconstruction of the past primarily relies on variation in present-day populations, limiting historical inference to lineages that are found among living subjects. Our analysis of four population groups in the Gaspé Peninsula, demonstrates how this may occasionally lead to erroneous interpretations. Mitochondrial DNA analysis of Gaspesians revealed an important admixture with Native Americans. The most likely scenario links this admixture to French-Canadians from the St. Lawrence Valley who moved to Gaspesia in the 19th century. However, in contrast to genetic data, analysis of genealogical record shows that Native American maternal lineages were brought to Gaspesia in the 18th century by Acadians who settled on the south-western coast of the peninsula. Intriguingly, within three generations, virtually all Métis Acadian families separated from their nonadmixed relatives and moved eastward mixing in with other Gaspesian groups, in which Native American maternal lines are present in relatively high frequencies. Over time, the carriers of these lines eventually lost memory of their mixed Amerindian-Acadian origin. Our results show that a reliable reconstruction of population history requires cross-verification of different data sources for consistency, thus favouring multidisciplinary approaches.