Archaeal Tuc1/Ncs6 homolog required for wobble uridine tRNA thiolation is associated with ubiquitin-proteasome, translation, and RNA processing system homologs.

While cytoplasmic tRNA 2-thiolation protein 1 (Tuc1/Ncs6) and ubiquitin-related modifier-1 (Urm1) are important in the 2-thiolation of 5-methoxycarbonylmethyl-2-thiouridine (mcm5s2U) at wobble uridines of tRNAs in eukaryotes, the biocatalytic roles and properties of Ncs6/Tuc1 and its homologs are poorly understood. Here we present the first report of an Ncs6 homolog of archaea (NcsA of Haloferax volcanii) that is essential for maintaining cellular pools of thiolated tRNA(Lys)UUU and for growth at high temperature. When purified from Hfx. volcanii, NcsA was found to be modified at Lys204 by isopeptide linkage to polymeric chains of the ubiquitin-fold protein SAMP2. The ubiquitin-activating E1 enzyme homolog of archaea (UbaA) was required for this covalent modification. Non-covalent protein partners that specifically associated with NcsA were also identified including UbaA, SAMP2, proteasome activating nucleotidase (PAN)-A/1, translation elongation factor aEF-1α and a ß-CASP ribonuclease homolog of the archaeal cleavage and polyadenylation specificity factor 1 family (aCPSF1). Together, our study reveals that NcsA is essential for growth at high temperature, required for formation of thiolated tRNA(Lys)UUU and intimately linked to homologs of ubiquitin-proteasome, translation and RNA processing systems.

Increased recognition of Powassan encephalitis in the United States, 1999-2005.

Powassan virus (POWV) disease is a rare human disease caused by a tick-borne encephalitis group flavivirus maintained in a transmission cycle between Ixodes cookei and other ixodid ticks and small and medium-sized mammals. During 1958-1998, only 27 POWV disease cases (mostly Powassan encephalitis) were reported from eastern Canada and the northeastern United States (average, 0.7 cases per year). During 1999-2005, nine cases (described herein) of serologically confirmed POWV disease were reported in the United States (average, 1.3 cases per year): four from Maine, two from New York, and one each from Michigan, Vermont, and Wisconsin. The Michigan and Wisconsin cases are the first ever reported from the north-central United States. Of these nine patients, 5 (56%) were men, the median age was 69 years (range: 25-91 years), and 6 (67%) had onset during May-July. All but one patient developed encephalitis with acute onset of profound muscle weakness, confusion, and other severe neurologic signs. In one case, no neurologic symptoms were present but the presence of pleocytosis, an elevated cerebrospinal fluid (CSF) protein concentration, and POWV-specific immunoglobulin M in CSF suggested neuroinvasion. All patients recovered from their acute disease, but most had long-term neurologic sequelae. Periresidential ecologic investigations were performed in three cases, including tests of local mammals and ticks for evidence of POWV infection. Woodchucks (Marmota monax), striped skunks (Mephitis mephitis), and a raccoon (Procyon lotor) collected at two of the Maine case-patients' residences had neutralizing antibody titers to POWV. I. cookei were found on woodchucks and skunks and questing in grassy areas of one of these residences; all were negative for POWV. Although POWV disease is rare, it is probably under-recognized, and it causes significant morbidity, and thus is an additional tick-borne emerging infectious disease entity. Because no vaccine or specific therapy is available, the basis of prevention is personal protection from ticks (or "tick hygiene") and reduced exposure to peridomestic wild mammals.

First U.S. record of the hard tick Ixodes (Pholeoixodes) gregsoni Lindquist, Wu, and Redner.

Ixodes (Pholeoixodes) gregsoni Lindquist, Wu, and Redner, a species of hard tick described in 1999 in Canada, was recovered from a harvested fisher (Martes pennanti Erxleben) and a domestic cat (Felis silvestris catus Ragni and Randi) in Vermont in 2001 and from harvested mink (Mustela vison Schreber) in Maine in 2003. These samples are the first records of this species within the United States. Although knowledge of this tick's natural history and distribution are still preliminary, these records indicate a possible greater distribution for I. gregsoni than initially perceived. Although its status as a disease vector is presently unknown, natural resource professionals should be aware of the possibility of this tick's occurrence in the northeastern United States.

Deer browse resistant exotic-invasive understory: an indicator of elevated human risk of exposure to Ixodes scapularis (Acari: Ixodidae) in southern coastal Maine woodlands.

We evaluated the relationships between forest understory structure and the abundance of questing adult and nymphal blacklegged ticks, Ixodes scapularis Say (Acari: Ixodidae), in three Maine towns endemic for Lyme disease, 2001-2003. In fragmented New England woodlands, over-abundant white-tailed deer, Odocoileus virginianus Zimmerman, overbrowse palatable species, allowing browse-resistant exotic-invasive species to replace native forest understory structures. We predicted there would be more ticks in plots dominated by exotic-invasive shrubs (such as Japanese barberry, Berberis thunbergii DC) than in plots dominated by native shrubs, ferns, or open understory. We assessed canopy composition and closure, tree basal area, litter composition, percentage of coverage and stem density of understory species, litter depth, soil moisture, and abundance of small mammals and white-tailed deer pellet groups. We used generalized linear mixed model analysis of covariance to determine the effect of understory structure on tick counts, controlling for continuous habitat and host covariates and adjusting for random spatial effects. There were twice as many adults and nearly twice as many nymphs in plots dominated by exotic-invasives than in plots dominated by native shrubs. Both adult and nymphal counts were lowest in open understory with coniferous litter. Adults were positively associated with increasing litter depth, medium soil moisture, and increasing abundance of white-footed deer mice, Peromyscus leucopus Rafinesque, and deer pellet group counts. Nymphs were positively associated with increasing litter depth, moderately wet soil, and mice. We concluded that deer browse-resistant exotic-invasive understory vegetation presented an elevated risk of human exposure to the vector tick of Lyme disease.

Macrophages and myeloid dendritic cells, but not plasmacytoid dendritic cells, produce IL-10 in response to MyD88- and TRIF-dependent TLR signals, and TLR-independent signals.

We have previously reported that mouse plasmacytoid dendritic cells (DC) produce high levels of IL-12p70, whereas bone marrow-derived myeloid DC and splenic DC produce substantially lower levels of this cytokine when activated with the TLR-9 ligand CpG. We now show that in response to CpG stimulation, high levels of IL-10 are secreted by macrophages, intermediate levels by myeloid DC, but no detectable IL-10 is secreted by plasmacytoid DC. MyD88-dependent TLR signals (TLR4, 7, 9 ligation), Toll/IL-1 receptor domain-containing adaptor-dependent TLR signals (TLR3, 4 ligation) as well as non-TLR signals (CD40 ligation) induced macrophages and myeloid DC to produce IL-10 in addition to proinflammatory cytokines. IL-12p70 expression in response to CpG was suppressed by endogenous IL-10 in macrophages, in myeloid DC, and to an even greater extent in splenic CD8alpha(-) and CD8alpha(+) DC. Although plasmacytoid DC did not produce IL-10 upon stimulation, addition of this cytokine exogenously suppressed their production of IL-12, TNF, and IFN-alpha, showing trans but not autocrine regulation of these cytokines by IL-10 in plasmacytoid DC.

A checklist of the mosquitoes of Maine with new state records.

A revised checklist of the mosquito fauna known to occur in Maine is reported. In addition we are detailing the finding of eight new state records in five genera, that is, Anopheles barberi, Culiseta minnesotae, Ochlerotatus dorsalis, Oc. japonicus, Oc. riparius, Oc. taeniorhynchus, Psorophora ferox, and Uranotaenia sapphirina. Locality records are given.

Abundance of Ixodes scapularis (Acari: Ixodidae) after the complete removal of deer from an isolated offshore island, endemic for Lyme Disease.

Monhegan is an isolated 237-ha island lying 16 km off the coast of Maine. Introduced to the island in 1955, white-tailed deer, Odocoileus virginianus Zimmerman, reached a density of approximately 37/km2 by the mid-1990s. Black-legged ticks, Ixodes scapularis Say, first noticed in the late 1980s, flourished thereafter. Norway rats (Rattus norvegicus Berkenhout) on Monhegan are highly infected with Borrelia burgdorferi Johnson, Schmidt, Hyde, Steigerwalt, and Brenner, the agent of Lyme disease. By 1996, 13% of year-round residents had contracted the disease. The community's subsequent decision to eliminate deer from the island provided a unique opportunity to monitor the abundance of vector ticks in response to the complete and permanent removal of the primary hosts of their reproductive stage. With the exception of humans and their dogs and cats, there are no other potential hosts for adult I. scapularis on Monhegan. From November 1996 to March 1999, all deer were removed from the island. Previous annual fall flagging of vegetation from 1990 to 1998 produced 6-17 adult ticks/h, of which 24-41% were infected with the Lyme disease spirochete. During this same period, up to 18 larvae and 4 nymphs were removed per Norway rat live-trapped on the island each July. With the absence of deer in the fall of 1999, both the density of host-seeking adult ticks and infection prevalence rose substantially to 28/h and 75.0%, respectively. By the summer of 2003, however, no sub-adult ticks were found on rats, and that fall, only 0.67 adult ticks/h were flagged. Of the 68 adults collected from 2002 to 2003, 20 (29.4%) were infected. Over this same period, adult tick abundance on a deer-populated, reference island continued to gradually increase.

Thermal accumulation and the early development of Ixodes scapularis.

We examined the relationship between the accumulation of thermal energy and the onset of oviposition and eclosion of the northern deer tick, Ixodes scapularis, and explored the usefulness of comparing degree days (DD) required for larval emergence with area-wide National Weather Service (NWS) data to construct maps indicating where the establishment of this vector tick would be climatologically constrained. Initially, the validity of basal temperatures for egg and larval development was confirmed by prolonged incubations of gravid females and eggs at 6 degrees C and 10 degrees C respectively. Next, the number of DD accumulated in situ from the placement of gravid females to oviposition, and from oviposition to larval emergence, were measured using temperature data loggers placed next to fall- and spring-fed ticks held within individual vials under leaf litter in multiple enclosures located in diverse biophysical regions of Maine. Finally, when it was found that total DD to larval emergence, as measured in ambient air above the enclosures, compared favorably with DD accumulated simultaneously at nearby NWS stations, maps were constructed, based on archived NWS data, to demonstrate where temperatures were sufficient to allow the hatching of larvae both within one season and over the last three decades as I. scapularis has advanced into northern New England.

Anaplasma phagocytophilum, Babesia microti, and Borrelia burgdorferi in Ixodes scapularis, southern coastal Maine.

Ixodes scapularis (deer ticks) from Maine were tested for multiple infections by polymerase chain reaction and immunofluorescence. In 1995, 29.5%, 9.5%, and 1.9% of deer ticks were infected with Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti, respectively. In 1996 and 1997, the number of A. phagocytophilum-infected ticks markedly declined. In 1995 through 1996, 4 (1.3%) of 301 were co-infected.

Deer density and the abundance of Ixodes scapularis (Acari: Ixodidae).

The abundance of Ixodes scapularis Say (Ixodes dammini Spielman, Clifford, Piesman & Corwin), the vector tick of the Lyme disease spirochete and other human pathogens, is related to the presence of its primary reproductive stage host, white-tailed deer (Odocoileus virginianus Zimmerman). However, this relationship has not been quantified in terms that would guide wildlife management in areas in which the public is, or is likely to become, exposed to infected ticks. In this study, deer density and tick abundance were measured in an emergent area for Lyme disease at three spatial scales using estimation methods appropriate for each. Simple linear regression was used to relate (1) the number of ticks found on deer at tagging stations in southern Maine to harvest-derived estimates of the density of deer within the towns in which they were killed, (2) tick densities estimated from fall flagging counts to deer densities estimated from pellet group counts made within multiple transects distributed through 5.2-km2 study sites, and (3) tick counts to pellet group counts within the individual transects. At the broadest scale, ticks on deer decreased with elevation and distance from the coast and increased with deer density, although deer and tick presence were only weakly related. Among the 5.2-km2 study sites and within individual transects, tick abundance related more strongly to deer pellet group counts. Few ticks were collected at deer densities <7/km2.

BCR targets cyclin D2 via Btk and the p85alpha subunit of PI3-K to induce cell cycle progression in primary mouse B cells.

The p85alpha subunit of PI3-K and Btk are two crucial components of the B-cell receptor (BCR) signalling pathway. In the present study, we showed that primary splenic B cells from p85alpha null and xid (Btk-deficient) mice fail to induce cyclin D2 expression and enter early G1, but not S phase of the cell cycle in response to BCR engagement. Furthermore, these Btk or p85alpha null B cells displayed increased cell death compared with wild type following BCR engagement. These findings are further confirmed by studies showing that specific pharmacological inhibitors of Btk (LFM-A13), PI3-K (LY294002 and Wortmannin) and PLCgamma (U73122) also block cyclin D2 expression and S phase entry following BCR stimulation, as well as triggering apoptosis. Collectively, these data provide evidence for the concept that the B-cell signalosome (p85alpha, Btk, BLNK and PLCgamma) is involved in regulating cyclin D2 expression in response to BCR engagement. PKC and intracellular calcium are two major downstream effectors of the B-cell signalosome and can be activated by PMA and ionomycin, respectively. In small resting (G0) B cells, costimulation with PMA and ionomycin, but not PMA or ionomycin alone, induces cyclin D2 expression and cell-cycle progression. Consistent with this, we also showed that the BCR-mediated cyclin D2 induction could be abolished by pretreatment of resting B cells with specific inhibitors of capacitative Ca(2+) entry (SK&F 96365) or PKC (Gö6850). Our present results lead us to propose a model in which the B-cell signalosome targets cyclin D2 via the Ca(2+) and PKC-dependent signalling cascades to mediate cell-cycle progression in response to BCR engagement.

Clinical characteristics and treatment outcome of early Lyme disease in patients with microbiologically confirmed erythema migrans.

BACKGROUND: Lyme disease has a wide spectrum of clinical manifestations. Diagnosis is usually based on the clinical and serologic picture rather than on microbiological confirmation. OBJECTIVE: To examine the clinical presentation and treatment outcome of early Lyme disease in patients with microbiologically confirmed erythema migrans. DESIGN: Observational cohort study. SETTING: 31 university-based or clinician-practice sites in 10 endemic states. PARTICIPANTS: 10 936 participants enrolled in a phase III trial of Lyme disease vaccine; 118 participants had erythema migrans in which Borrelia burgdorferi was detected by culture or polymerase chain reaction. MEASUREMENTS: Clinical characteristics and treatment outcome were noted. Skin biopsies of erythema migrans were performed for culture and detection of B. burgdorferi by polymerase chain reaction; serologic responses were determined by Western blot. RESULTS: The 118 patients with microbiologically confirmed erythema migrans presented a median of 3 days after symptom onset. Early erythema migrans commonly had homogeneous or central redness rather than a peripheral erythema with partial central clearing. The most common associated symptoms were low-grade fever, headache, neck stiffness, arthralgia, myalgia, or fatigue. By convalescence, 65% of patients had positive IgM or IgG antibody responses to B. burgdorferi. Most patients responded promptly to antibiotic treatment. CONCLUSIONS: In major endemic areas in the United States, Lyme disease commonly presents as erythema migrans with homogeneous or central redness and nonspecific flu-like symptoms. Clinical outcome is excellent if antibiotic therapy is administered soon after symptom onset.

Bottomley is still causing damage.

Poor Mrs Bottomley, shuffled away to Heritage but still managing to scratch at the health of our nation.