Characterization of a novel staphylococcal cassette chromosome composite island from community-associated MRSA isolated in aged care facilities in Western Australia.

BACKGROUND: In Western Australia (WA), clonal complex 5, ST835, community-associated (CA) MRSA is isolated almost exclusively from aged care facilities. In WA four different staphylococcal cassette chromosome (SCC) mec (SCCmec) elements have been identified in this ST, indicating high genetic activity in the SCCmec region. OBJECTIVES: To investigate the SCC region of ST835 CA-MRSA WA MRSA-40 and determine the distribution of an SCCsorbitol element found within the region. RESULTS: The SCC region contained a composite island, SCCmecWA MRSA-40-CI, that was composed of three elements, ΨSCCpls, SCCsorbitol and SCCmecVT (5C2&5). This is the first time that a sorbitol operon has been reported in an SCC element. CONCLUSIONS: Generation of SCCmecWA MRSA-40-CI has involved multiple genetic events and recombination with CoNS has occurred during evolution of the SCC elements. While Staphylococcus aureus is renowned for its ability to utilize mobile genetic elements to disseminate antimicrobial resistance, the SCC region of WA MRSA-40 shows that this clone has also utilized SCC elements to acquire extra virulence and possibly adapt to a niche environment.

Spontaneous limb movements and posturing secondary to acute basilar artery occlusion: a potentially devastating seizure mimic.

Basilar artery occlusion is a devastating but treatable form of ischaemic stroke with high morbidity and mortality rates. The diagnosis is often challenging due to considerable heterogeneity of clinical signs and symptoms. We report a case of an acute basilar artery occlusion presenting with decreased level of consciousness associated with rhythmic tonic movements of the four extremities, mimicking seizure activity. The patient was treated with intravenous thrombolysis and subsequently gained good recovery. Awareness of this entity is required to recognise this potentially treatable, but otherwise devastating seizure mimic.

Properties of blood, porphyrins, and exposure to legacy and emerging persistent organic pollutants in surf scoters (Melanitta perspicillata) overwintering on the south coast of British Columbia, Canada.

The surf scoter (Melanitta perspicillata) is a little-studied species of North American sea duck. Estimates suggest it has experienced a precipitous decline in breeding numbers over the latter half of the past century. To investigate the potential role of contaminant uptake and toxicity in the population decline, this study undertook to measure blood chemistry, porphyrin concentrations, EROD, and organic contaminants in mature surf scoters wintering in the Strait of Georgia, BC, Canada. Hepatic organochlorine pesticide, polychlorinated dibenzo-p-dioxin, polychlorinated dibenzofuran, polychlorinated biphenyl (PCB), polybrominated diphenyl ether, and nonylphenol concentrations were relatively low; for example, SigmaTEQs (toxic equivalents) for PCBs, dioxins, and furans combined ranged from 4.7 ng/kg wet weight in reference-site (Baynes Sound) birds to 11.4 ng/kg wet weight in birds from Vancouver Harbour. Nonetheless, elevated EROD activity indicated that birds in Howe Sound were responding to an Ah-receptor-mediated stressor, which was also affecting hematocrit values and possibly vitamin A status. In addition, a low proportion of lymphocytes in individuals across locations in early spring samples was associated with poor body condition. The apparent loss of fitness just prior to the onset of northerly migrations to breeding grounds is of particular concern. Compromised health of mature birds at this point in the season might impact negatively on the productivity and survival of some individuals, particularly those overwintering in Howe Sound.

The outcome of non-typhoidal salmonella meningitis in Malawian children, 1997-2006.

INTRODUCTION: The clinical course and outcome of non-typhoidal salmonella (NTS) meningitis in Malawian children over a 10-year period (1997-2006) is described. METHODS: Demographic, clinical and laboratory data were collected for all children over 2 months of age admitted with salmonella meningitis to Queen Elizabeth Central Hospital from 1997 to 2006. In the 1st year, salmonellae were susceptible to chloramphenicol, and children received 2 weeks of chloramphenicol treatment. When NTS resistance to chloramphenicol started to appear in 1998, treatment was changed to ceftriaxone. From 2002, the duration of antibiotic therapy was extended to 4-weeks which included 2 weeks of intravenous ceftriaxone and a further 2 weeks of oral ciprofloxacin. RESULTS: The in-hospital case fatality rate (CFR) was 52.3% (48.2% until 2002 and 53.9% after prolonged antibiotic therapy was introduced). Of the survivors, one in 12 (8.3%) became completely well (sequelae-free) in the period 1997-2001 while 18 of 31 survivors (58.1%) made a complete recovery during 2002-2006 (p<0.01). After the 4-week course of antimicrobial therapy was introduced, the number of relapses or recurrences fell from nine in 15 (60%) survivors treated with chloramphenicol or ceftriaxone to three in 35 (8.7%) survivors who received 4 weeks of antibiotics (p<0.0001). CONCLUSION: In Malawi, salmonella meningitis has a CFR of approximately 50%, which has remained constant over many years. Residual morbidity, however, has decreased over 10 years, despite rising numbers of multi-drug-resistant cases of NTS. This improvement might be owing to better treatment and management and/or reduced pathogenicity of the multi-drug-resistant bacteria.

Butyltins, trace metals and morphological variables in surf scoter (Melanitta perspicillata) wintering on the south coast of British Columbia, Canada.

From 1998 to 2001 we examined spatial and temporal variation in uptake of contaminants by surf scoters (Melanitta perspicillata) in the Georgia Basin region of the Pacific coast of Canada. Samples were collected during late fall and early spring at industrialized and reference locations, carcasses examined, and tissues collected for histology, biomarkers, and contaminant analyses. Scoters from both Vancouver and Victoria harbours had significantly higher hepatic concentrations of summation operatorbutyltins than birds from a reference site. In adult male surf scoters, hepatic summation operatorbutyltins increased over the winter at two sites (p=0.02, n=26), while mercury increased (p=0.03, n=15) and selenium decreased at one site (p=0.001, n=15). Body condition decreased over the winter at both the treatment site, Howe Sound (p<0.0001, n=12), and the reference site, Baynes Sound (p=0.02, n=15). Multiple regression analysis using Akaike's Information Criteria (AIC(C)) showed an association between hepatic butyltin concentrations and overall body condition (p=0.06, r=-0.237).

Vitamin A and contaminant concentrations in surf scoters (Melanitta perspicillata) wintering on the Pacific coast of British Columbia, Canada.

Surf scoters are part of a community of sea ducks on the western coast of North America that have shown signs of long-term, unexplained declines in breeding bird numbers. Substantial numbers of scoters winter in the major harbours on the west coast, after breeding in the west-central northern boreal forest. To address the potential for contaminants to impact the health and survival of those birds, we investigated the condition and contamination of surf scoters during the winters of 1998-2001 at four foraging locations in the Strait of Georgia region of the Pacific coast of Canada. Vitamin A status was evaluated in liver and plasma samples collected from adults and juveniles, as part of a larger assessment of tissue contamination, body condition and biomarker responses. Individuals collected from a relatively contaminated site, Howe Sound, showed consistently low hepatic concentrations of retinol and retinyl palmitate forms of vitamin A, and gender-specific associations of retinyl palmitate with hepatic EROD activity. The relationship of hepatic retinol to retinyl palmitate was not constant across geographic locations, and a clear, linear relationship between the two forms of vitamin A was only evident in birds from the relatively uncontaminated site. This study also identified strong positive relationships between vitamin A and tissue burdens of cadmium and zinc. The positive association between hepatic retinyl palmitate and renal cadmium is similar to one observed in laboratory rats, in which a mechanism of interference with the controlled release of retinol from the liver was suggested.

Aetiology of neonatal sepsis in Blantyre, Malawi: 1996-2001.

AIM: The aim of this retrospective study was to report causes, antibiotic resistance and outcome of neonatal sepsis (often fatal in developing countries) in Malawi. METHODS: All blood and cerebrospinal fluid isolates collected between January 1996 and December 2001 from inpatients aged 0-30 days with suspected sepsis at Queen Elizabeth Central Hospital, Blantyre, Malawi were reviewed. In vitro resistance to antibiotics commonly used in Malawi was assessed. Case fatality rate was analysed with respect to age, bacterial pathogen and infection site. RESULTS: A total of 801 bacteria were isolated from 784 neonates over 6 years-599 isolates from blood and 202 from cerebrospinal fluid. Overall, 54% of bacteria were gram-positive and 46% gram-negative. The commonest causes of neonatal sepsis were group B Streptococcus (17%) and non-typhoidal Salmonella (14%). In vitro antibiotic susceptibility to the first-line antibiotic combination of penicillin and gentamicin was 78% for all isolates, but in vitro sensitivities to gentamicin for Klebsiella spp and non-typhoidal Salmonella were only 33% and 53%, respectively. In-hospital case fatality rate was known for only 301 cases and was high at 48%. Group B Streptococcus was associated with the best outcome. Mortality was significantly higher if presentation was in the 1st week of life or if sepsis was caused by gram-negative bacteria. The causes of neonatal sepsis in this population show a different pattern from other studies in developing countries.

Assessing breeding potential of peregrine falcons based on chlorinated hydrocarbon concentrations in prey.

Peregrine falcons (Falco peregrinus) now breed successfully in most areas of North America from which they were previously extirpated. The loss during the mid-part of the last century of many of the world's peregrine populations was largely a consequence of impaired reproduction caused by the effects of DDE on eggshell quality and embryo hatchability. Population recovery has been attributed to re-introduction efforts, coupled with regulatory restrictions on the use of organochlorine pesticides. Peregrines have not returned to breed in some areas, such as the Okanagan Valley of British Columbia. That region has been extensively planted in fruit orchards which were treated annually with DDT during the early 1950s to the 1970s. Ongoing contamination of avian species, including potential peregrine prey, inhabiting orchards has been documented. In response to an initiative to release peregrines around the city of Kelowna in the Okanagan Valley, we collected potential peregrine prey species and analyzed whole bodies for chlorinated hydrocarbon residues. We used a simple bioaccumulation model to predict concentrations of DDE in peregrine eggs using concentrations in prey and estimates of dietary makeup as input. Peregrines would be expected to breed successfully only if they fed on a diet primarily of doves. Feeding on as little as 10% of other species such as starlings, robins, gulls and magpies would produce DDE concentrations in peregrine eggs greater than the threshold of 15 mg/kg. We also estimated the critical concentration of DDE in total prey to be about 0.5 mg/kg, one half of the previous most conservative criterion for peregrine prey. Concentrations of dieldrin and PCBs in peregrine prey are less than suggested critical levels.

Reproductive success and chlorinated hydrocarbon contamination of resident great blue herons (Ardea herodias) from coastal British Columbia, Canada, 1977 to 2000.

Over the period 1977-2000, eggs of Pacific great blue heron (Ardea herodias fannini) were collected from 23 colonies along the southern coast of British Columbia, Canada, and analyzed for persistent organochlorine (OC) pesticides and polychlorinated biphenyls (PCBs). Concentrations of OC pesticides in eggs declined sharply in the late 1970s, after which there were minimal changes. The sums of PCB congeners were not reduced appreciably during the 1980s and 1990s, but Aroclor 1260 concentrations suggested a sharp decline in PCB contamination of eggs in the late 1970s, similar to that shown for OC pesticides. Eggs collected along or near the Fraser River delta showed higher levels of most pesticides compared to other monitored colonies. Although the delta lands support a long-standing agricultural economy, the primary factors influencing OC levels in the delta colonies were thought to be driven by estuarine processes. We suggest two possible influencing factors were: 1) a greater rate of bioaccumulation in the estuary due to the deposition of particulates collected over a vast area encompassed by the Fraser River watershed; or 2) a higher rate of biomagnification in the estuary due to species differences at lower trophic levels of the heron food chain. Eggs from urban colonies contained higher levels of PCBs. The congener pattern was not clearly different from that observed in less contaminated eggs from rural and pulp mill-influenced colonies, except that colonies in Vancouver had greater proportions of PCB-66, suggesting a local source of Aroclor 1242. Productivity in the coastal heron colonies was highly variable over the period of study, with 71% of recorded colony-wide reproductive failures occurring in colonies near pulp mills. However, the predominant factors influencing reproductive success were probably disturbance by humans and bald eagles, combined with loss and degradation of nesting habitat, and not sublethal toxicity.

Surveillance of invasive infection in children and adults admitted to QECH, Blantyre, 1996-2002.

This is a report of blood CSF isolates from the adults medical and paediatric of wards QECH, Blantyre, cultured and identified at the Welcome Trust Research Laboratories during 1996-2002. The commonest causes of adults and children bacteraemia were non-typhoidal Salmonella (35% of all blood isolates for adults and children) and Streptococcus pneumoniae (14% and 13% respectively). Cryptococcus neoformans was the commonest isolates from CSF of adults with meningitis(67%) but was very rare in children. S.pneumoniae was the commonest cause of bacterial meningitis in children and adults (38% and 28% of all CSF isolates respectively). Haemophilus influenzae type b was also a common cause of meningitis in children (27%). Data of in vitro antibiotic sensitivity are also reported. A major concern is the recent marked rise of chloramphenicol resistance among Salmonella enteritidis and Salmonella typhimurium to over 80% resistance.

Mercury residues in livers of bald eagles (Haliaeetus leucocephalus) found dead or dying in British Columbia, Canada (1987-1994).

Postmortem examinations were conducted on 82 bald eagles (Haliaeetus leucocephalus) found dead or dying in British Columbia, Canada, from 1987 to 1994. As part of the examination, livers were analyzed for total mercury (Hg) content, as well as methylmercury (meHg) and selenium (Se) in selected individuals. In total, 67 eagles were classed as having low Hg exposure [total Hg liver residues ranging from 0.5 to 17.2 mg/kg dry weight (dw)]. Fourteen eagles were moderately exposed (liver residues ranging from 19.2 to 36.8 mg/kg Hg dw). One eagle was judged to have died of Hg poisoning, with a total liver Hg content of 130.3 mg/kg dw, of which approximately 77% was meHg. The poisoned eagle and most of the exposed eagles were found in locations where effluent from pulp and paper processing plants is discharged along the British Columbia coast. In total, 6% of eagles examined died as a result of acute metal toxicosis (one from Hg poisoning, four from lead poisoning), in comparison to 72% dying from trauma (electrocution, vehicle/power line collision, eagle attack, trap, gunshot, drowning, and asphyxiation) and 11% from disease. The cause of death was undetermined in the remaining 11% of eagles.

Assessment of biological effects of chlorinated hydrocarbons in osprey chicks.

Osprey (Pandion haliaetus) eggs were collected during 1995 and 1996 at seven sites along the Fraser and Columbia River systems of British Columbia, Canada, and Washington and Oregon, USA. Fifty-four eggs were placed into a laboratory incubator. Thirty-eight of the hatched chicks were sacrificed within 24 h. Hatching success did not differ among sites and therefore between treatment and reference areas. Residual yolk sacs of eggs collected downstream of the large bleached-kraft pulp mill at Castlegar contained greater mean concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, 2,930 ng/kg lipid) compared with reference sites such as the Nechako River, an upper tributary of the Fraser system (33.7 ng/kg). Total polychlorinated biphenyls (PCBs) in yolk sacs were also higher at Castlegar and in samples from the Columbia River downstream of Portland, Oregon, compared with those from the Nechako River. Concentrations of measured chemicals, including TCDD toxic equivalents (TEQs), total PCBs, p,p'-dichlorodiphenylethylene (p,p'-DDE), and other organochlorines were not different in eggs that failed to hatch compared with calculated whole-egg values for hatched eggs. There were significant biochemical responses; a hepatic cytochrome P4501A (CYP1A) cross-reactive protein was detected in all samples tested and correlated positively with ethoxyresorufin o-deethylase (EROD) activity and yolk sac concentrations of TEQs and total PCBs. Tissue concentrations of vitamin A compounds varied among sites and correlated positively with yolk sac concentrations of TEQs and PCBs. Morphological, histological, and other physiological parameters, including chick growth, edema, deformities, and hepatic and renal porphyrin concentrations, neither varied among sites nor showed concentration-related effects.

Monitoring temporal and spatial trends in polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in eggs of great blue heron (Ardea herodias) on the coast of British Columbia, Canada, 1983-1998.

Eggs from 21 resident great blue heron (Ardea herodias) rookeries were monitored from 1983 to 1998 along the coast of British Columbia, Canada, for contamination with polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and biphenyls (PCBs). Dominant congeners (1,2,3,7,8-PnCDD, 1,2,3,6,7,8-HxCDD, 2,3,7,8-TCDD, 2,3,7,8-TCDF, 1,2,3,7,8,9-HxCDD and 2,3,4,7,8-PnCDF) fell markedly in the early 1990s after pulp mills changed from molecular chlorine bleaching to alternative bleaching technologies, and the use of chlorophenolic wood preservatives and anti-sapstains was severely restricted. Strong positive linear regressions between prey fish and heron egg contaminant levels suggested that local dietary uptake was an important route of exposure for herons. Toxic equivalent concentrations (TEQs) sufficient to produce embryotoxicity in great blue heron chicks were measured in eggs from 1985 to 1991 at some colonies. Despite reduction in PCDD/Fs, estimated TEQs remained elevated throughout the 1980s at some urban colonies due to contributions from PCBs.

Transfer of DDT and metabolites from fruit orchard soils to American robins (Turdus migratorius) twenty years after agricultural use of DDT in Canada.

Wildlife contamination studies found high levels of DDT and associated metabolites in bird eggs from Canadian orchard sites during the early 1990s. The present study investigated local dietary uptake of DDT and geographic variability in tissue concentrations in the same orchards. A soil-earthworm-robin food chain was chosen for study, as early surveys showed that robins contained the highest levels of DDT of several avian species and because published research indicated that earthworms were a probable dietary exposure route. Organochlorine pesticides and PCBs were measured in soil, earthworm, robin egg, and robin nestling samples collected from fruit orchards and reference sites. High average DDE (soil: 5.2 mg/kg; earthworm: 52 mg/kg; robin egg: 484 mg/kg dry weight) and DDT (soil: 9.2 mg/kg; earthworm: 21 mg/kg; robin egg: 73 mg/kg dry weight) concentrations in Okanagan (British Columbia) samples confirmed that previously recorded contamination was common in the region. Concentrations detected in Simcoe, Ontario, orchards were not as high but were still significantly elevated relative to levels in soils and robins from reference areas. Significant positive linear regressions between soil and earthworm concentrations and consistent trends in food chain accumulation suggested that robins were acquiring DDT and metabolite (DDTr) burdens locally. Low concentrations of DDT and DDTr in robin eggs collected from nests in nearby nonorchard and post-DDT orchard habitats suggested that the local sources were in orchards. Persistence of DDT in orchard food chains is likely due to a combination of retarded degradation rates for DDT in soil and its extensive use historically. DDT concentrations in some robin eggs and earthworms were at levels comparable to those observed in field studies where mortality or reproductive effects occurred.

A complete system for identifying inhibitors of creatine kinase B.

We have developed a complete system for discovery of lead compounds as inhibitors of creatine kinase B. In this article, we describe production and purification of the recombinant protein, conditions and features of an optimized high-throughput screening assay, and results of our implementation of the system using a diverse compound library.

Contaminants in ospreys from the Pacific Northwest: II. Organochlorine pesticides, polychlorinated biphenyls, and mercury, 1991-1997.

Eggs of ospreys (Pandion haliaetus) were collected over the period 1991 to 1997 at 111 nests in the Fraser and Columbia River drainage systems of British Columbia, Washington, and Oregon. Contents were analyzed for organochlorine (OC) pesticides, polychlorinated biphenyls (PCBs), and total mercury. Blood samples were taken from nestling ospreys at two time points during the 1992 breeding season on the Thompson River and analyzed for non-ortho PCBs. Concentrations of DDE and related compounds showed high variability among individual eggs within study areas and no significant differences in mean concentrations among study areas. Some eggs contained high concentrations of DDE, up to 20 mg/kg (wet weight), for which there were no evident local sources, suggesting that exposure occurred outside of the breeding grounds. Most other OC pesticides measured in osprey eggs showed a similar distribution, although mean concentrations of chlordane-related compounds were generally, and in some cases significantly, higher in samples collected from the Columbia River sites compared to elsewhere. Significantly greater concentrations of PCBs were found in eggs from the Columbia River basin compared to the Fraser. This was attributed to extensive development of hydroelectric generation and related industries on the Columbia system. Significant differences among sites were also found in the pattern of PCB congeners, e.g., eggs from the lower Columbia River site had proportionally greater concentrations of less chlorinated, Aroclor 1242-type PCBs compared to other sites. In contrast, eggs from upper reaches of the Columbia River had relatively greater amounts of Aroclor 1254 and 1260 congeners. Mercury concentrations in osprey eggs tended to be uniform among sites and comparable to those reported in the literature for ospreys nesting on naturally formed lakes and rivers. There were no significant temporal changes in mean concentrations of any of the measured compounds at sites monitored from 1991 to 1997.

Prediction of serum IgG1 concentration in beef calves based on age and serum gamma-glutamyl-transferase activity.

This study examined the relationship between serum gamma-glutamyl-transferase (GGT) activity and passive transfer status in beef calves less than 18 days of age. Immunoglobulin G1 (IgG1) concentrations were measured in 69 commercial beef calves between the ages of 24 and 72 hours. GGT activities were then measured in these same calves at various ages between 3 and 18 days of age. Models were developed predicting serum IgG1 concentration as a function of calf age and serum GGT activity. Minimal association was present between initial serum IgG1 concentration and serum GGT activity when all calves less than 18 days of age were considered (r2 = 0.065). When the study population was restricted to calves less than 8 days of age, the fit of the developed model was greatly improved (r2 = 0.438). Serum GGT activity has no apparent advantage relative to other assay procedures for predicting passive transfer status in beef calves. If serum GGT activity is to be used to assess passive transfer status in beef calves, application of this procedure should be restricted to calves less than 8 days of age.

Contaminants in ospreys from the Pacific Northwest: I. Trends and patterns in polychlorinated dibenzo-p-dioxins and -dibenzofurans in eggs and plasma.

Osprey (Pandion haliaetus) eggs were collected from 1991 to 1997 at nests (n = 121) upstream and downstream of bleached kraft pulp mills and at reference sites in the Fraser and Columbia River drainage systems of British Columbia, Washington, and Oregon. Blood samples were collected from nestling ospreys during the 1992 breeding season on the Thompson River. Samples were analyzed for polychlorinated dibenzo-p-dioxins (PCDDs) and -dibenzofurans (PCDFs). Mean concentrations of 2,3,7,8-TCDD were significantly higher in eggs collected in 1991 at downstream compared to upstream nests near pulp mills at Kamloops and Castlegar, British Columbia. There were no significant temporal trends in 2,3,7,8-TCDD, -TCDF or other measured compounds at a sample of nests monitored between 1991 and 1994 downstream of the Castlegar pulp mill, despite changes in bleaching technology (CIO2 substitution). However, by 1997 concentrations of 2, 3,7,8-TCDD and -TCDF were significantly lower than previous years in nests sampled downstream at both Castlegar and Kamloops. An unusual pattern of higher chlorinated PCDDs and PCDFs was found in many of the osprey eggs collected in this study, and considerable individual variation in the pattern existed among eggs from the same site. For example, eggs from four different nests at one study area (Quesnel) on the Fraser River had concentrations of 1,2,3,4,6,7,8-HpCDD ranging from <1 to 1,100 ng/kg and OCDD from <1 to 7,000 ng/kg wet weight. Higher mean concentrations of HpCDD and OCDD were found in eggs from the Thompson River, a tributary of the Fraser, compared to the Columbia River, and concentrations were generally higher at nests upstream of pulp mills. In plasma samples, 1,2,3,4,6,7,8-HpCDD and OCDD were the main compounds detected, with no significant differences measured between samples upstream versus downstream or earlier versus later in the breeding season. Use of chlorophenolic wood preservatives by lumber processors was considered the main source of higher chlorinated PCDD/Fs throughout the systems, based on patterns of trace PCDFs in eggs and significant correlations between egg concentrations of pentachlorophenol and both HpCDD (r = 0.891, p < 0.01) and OCDD (r = 0.870, p < 0.01).

Casein kinase II catalyzes tyrosine phosphorylation of the yeast nucleolar immunophilin Fpr3.

In the yeast Saccharomyces cerevisiae, the nucleolar immunophilin, Fpr3, is phosphorylated at tyrosine and dephosphorylated by the phosphotyrosine-specific phosphoprotein phosphatase, Ptp1. In Ptp1-deficient cells, Fpr3 contains phospho-Tyr at a single site (Tyr184), but also contains phospho-Ser and phospho-Thr. Ser186 (adjacent to Tyr184) is situated within a canonical site for phosphorylation by casein kinase II (CKII). Yeast cell lysates contain an activity that binds to Fpr3 in vitro and phosphorylates Fpr3 at Ser, Thr, and Tyr; this activity was found to be dependent on expression of functional yeast CKII. Moreover, purified Fpr3 was phosphorylated on Tyr184 in vitro by either purified yeast CKII or purified, bacterially-expressed human CKII. Likewise, phosphorylation of Fpr3 at tyrosine in vivo was markedly enhanced in yeast cells overexpressing a heterologous (Drosophila) CKII, but was undetectable in yeast cells carrying only a temperature-sensitive allele of the endogenous CKII, even when the cells were grown at a permissive temperature. Phosphorylation of Fpr3 at Tyr184 by CKII in vitro lagged behind phosphorylation of Fpr3 at Ser, and was accelerated by pre-phosphorylation of Fpr3 at Ser using CKII. Furthermore, synthetic peptides corresponding to the sequence surrounding Tyr184 that contained P-Ser (or Glu) at position 186 were much more efficient substrates for CKII phosphorylation of Tyr184 than a synthetic peptide containing Ala at position 186. These findings indicate that CKII phosphorylates Fpr3 at tyrosine and serine both in vivo and in vitro and thus possesses dual specificity. These results also indicate that Tyr184 is phosphorylated by CKII via a two-step process, in which phosphorylation at the +2 position provides a negatively-charged specificity determinant that allows subsequent phosphorylation of Tyr184.