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Biomedical subjects

J F Anderson

Publications and source records attributed to J F Anderson.

At least 19 recordsLinked to original sources

A novel family of anticoagulants from the saliva of Ixodes scapularis.

Using biochemical and molecular approaches, we have identified a 9.8 kDa protein in the saliva of Ixodes scapularis that inhibits the intrinsic pathway of coagulation. The 9.8 kDa anticoagulant protein was purified by reverse-phase HPLC and its N-terminal amino acid sequence determined. The N-terminal sequence showed homology with Salp14, an immuno-dominant antigen present in the saliva of engorging I. scapularis nymphs. Recombinant Salp14 expressed in Escherichia coli prolonged the activated partial thromboplastin time (APTT) of human plasma in a dose-dependent manner and was a specific inhibitor of factor Xa. A cDNA encoding a 9.3 kDa protein, Salp9Pac, was subsequently isolated from an I. scapularis salivary gland cDNA library. Salp9Pac showed 93% identity to the N-terminal sequence of the anticoagulant purified by HPLC. These data indicate that the anticoagulant protein purified by HPLC, Salp9Pac and Salp14 are members of a family of novel coagulation protease inhibitors present in tick saliva. While recombinant Salp9Pac did not show biological activity in the assays tested currently, it is likely to be mechanistically different from its paralogues. This raises the possibility that ticks may enhance their adaptive ability to cope with a wide spectrum of proteases, by transcribing such structurally related anticoagulant proteins with different functions.

Amino Acid Sequence↗

Immunization of mice against West Nile virus with recombinant envelope protein.

West Nile (WN) virus is a mosquito-borne flavivirus that emerged in the United States in 1999 and can cause fatal encephalitis. Envelope (E) protein cDNA from a WN virus isolate recovered from Culex pipiens in Connecticut was expressed in Escherichia coli. The recombinant E protein was purified and used as Ag in immunoblot assays and immunization experiments. Patients with WN virus infection had Abs that recognized the recombinant E protein. C3H/HeN mice immunized with E protein developed E protein Abs and were protected from infection with WN virus. Passive administration of E protein antisera was also sufficient to afford immunity. E protein is a candidate vaccine to prevent WN virus infection.

Animals↗

A phylogenetic approach to following West Nile virus in Connecticut.

The 1999 outbreak of West Nile (WN) virus in the northeastern United States was the first known natural occurrence of this flavivirus in the Western Hemisphere. In 1999 and 2000, 82 independent Connecticut WN virus isolates were cultured from nine species of birds, five species of mosquitoes, and one striped skunk. Nucleotide sequences obtained from these isolates identified 30 genetic changes, compared with WN-NY99, in a 921-nt region of the viral genome beginning at nucleotide position 205 and ending at 1125. This region encodes portions of the nucleocapsid and envelope proteins and includes the entire coding regions for the premembrane and membrane proteins. Amino acid changes occurred at seven loci in six isolates relative to the WN-NY99 strain. Although 34 of the isolates showed sequences identical to the WN-NY99 isolate, we were able to show geographical-based clusters of mutations. In particular, 26 isolates were characterized by mutation of C to T at position 858. This group apparently originated in Stamford, CT and disseminated to sites located as far as 54 miles from Stamford. Sequences of WN virus isolated from both brain and heart tissues from the same avian host were identical in all 14 tested individual birds, suggesting that the mutations we have documented are real and not caused by culture, RNA extraction, or PCR procedures. We conclude that this portion of the viral genome will enable us to follow the geographical and temporal movement of variant WN virus strains as they adapt to North America.

Animals↗

Discovery, distribution, and abundance of the newly introduced mosquito Ochlerotatus japonicus (Diptera: Culicidae) in Connecticut, USA.

The earliest documented specimen of an exotic east Asian mosquito Ochlerotatus (Finlaya) japonicis japonicus (Theobald) in the Western Hemisphere is reported along with the results of a state wide survey to determine the distribution and abundance of this mosquito in Connecticut. Ochlerotatus japonicus was collected from 87 locations in eight counties. It is established throughout the state and occurs in a variety of natural and artificial container habitats including discarded tire casings, bird baths, wooden barrels, porcelain bath tubs (used for watering animals), plastic milk cartons, toys, vinyl tarpaulins (covering wood piles and swimming pools), exposed rock holes in stream beds, tree holes, subterranean catch basins, surface water rain pools, and spring-fed depressions. Larvae were particularly common in containers with water, decaying leaves, and algae, in shaded and sunlit areas and, in rock-pool habitats along streambeds, in association with Ochlerotatus atropalpus (Coquillett). Adult females were collected in sod grass-infused gravid and CO2- baited light traps, from early June through October, with peak collections in September. Biting females were collected by human bait method augmented with CO2, verifying its capacity to feed on humans. The ovitraps used in this study were not effective for recovering this species. Our results suggest that Oc. japonicus was introduced into Connecticut between 1992 and 1998. Because of the ability of Oc. japonicus to transmit West Nile virus, and because of the recent detection of this virus in field-collected specimens, the introduction of Oc. japonicus is considered a significant public health development.

Animals↗

West Nile virus envelope protein: role in diagnosis and immunity.

The role of antibodies to the West Nile virus envelope (E) protein in serodiagnosis and protection was examined. The E protein was expressed and purified in recombinant form. Antibodies to the E protein were detected in patients with West Nile virus infection. Passive immunization with rabbit anti-E protein sera also partially protected mice from challenge with West Nile virus. The humoral response to the West Nile virus E protein is therefore useful as an aid in the diagnosis and may also play a role in immunity to infection.

Animals↗

Hamular frenum modification: a removable denture prosthesis retention and stability enhancement.

A removable denture prosthesis, whether partial or complete, often requires preprosthetic surgery to achieve optimum stabilization and retention. While the hamular frenum may produce significant dynamic dislodging forces, a literature review did not reveal any reports dealing with this problem. A hamular frenum reduction surgical procedure using the free autogenous gingival graft procedure is described. Prosthetic function may be enhanced by eliminating the dynamic disrupting force of the hamular frenum along with improving posterior maxillary tuberosity contour and, as necessary, premaxillary form, allowing these contours to work in concert to develop a "cupping" stabilizing and retentive complex.

Aged↗

Detection of Ehrlichia chaffeensis DNA in Amblyomma americanum ticks in Connecticut and Rhode Island.

Ehrlichia chaffeensis, the causative agent of human monocytic ehrlichiosis, is transmitted by Amblyomma americanum ticks, which are most abundant in the southern United States. Because serologic evidence suggests that residents of Connecticut are exposed to E. chaffeensis, A. americanum ticks were collected in Connecticut and Rhode Island for PCR analysis to detect E. chaffeensis DNA. Eight of 106 (7.6%) A. americanum ticks from Connecticut and 6 of 52 (11.5%) from Rhode Island contained E. chaffeensis DNA. Thus, E. chaffeensis is present in ticks in southern New England and transmission of E. chaffeensis may occur there.

Animals↗

Recovery and identification of West Nile virus from a hawk in winter.

West Nile virus was recovered from the brain of a red-tailed hawk that died in Westchester County, N.Y., in February 2000. Multiple foci of glial cells, lymphocytes, and a few pyknotic nuclei were observed in the brain. Three to 4 days after inoculation of Vero cells with brain homogenates, cytopathic changes were detected. The presence of West Nile virus antigen in fixed cells or cell lysates was revealed by fluorescent antibody testing or enzyme-linked immunosorbent assay, respectively. Furthermore, Reverse transcriptase-PCR with primers specific for the NS3 gene of West Nile virus resulted in an amplicon of the expected size (470 bp). Electron microscopy of thin sections of infected Vero cells revealed the presence of viral particles approximately 40 nm in diameter, within cytoplasmic vesicles. The demonstration of infection with the West Nile virus in the dead of the winter, long after mosquitoes ceased to be active, is significant in that it testifies to the survival of the virus in the region beyond mosquito season and suggests another route of transmission: in this case, prey to predator.

Animals↗

Attachment of Borrelia burgdorferi within Ixodes scapularis mediated by outer surface protein A.

Borrelia burgdorferi outer surface protein (Osp) A has been used as a Lyme disease vaccine that blocks transmission: OspA antibodies of immune hosts enter ticks during blood feeding and destroy spirochetes before transmission to the host can occur. B. burgdorferi produce OspA in the gut of unfed Ixodes scapularis ticks, and many spirochetes repress OspA production during the feeding process. This preferential expression suggests that OspA may have an important function in the vector. Here we show that OspA mediates spirochete attachment to the tick gut by binding to an I. scapularis protein. The binding domains reside in the central region and COOH-terminus of OspA. OspA also binds to itself, suggesting that spirochete-spirochete interactions may further facilitate adherence in the gut. OspA-mediated attachment in the tick provides a possible mechanism for how stage-specific protein expression can contribute to pathogenesis during the B. burgdorferi natural cycle.

Amino Acid Sequence↗

Utilization of common analgesic and anxiolytic medications by registered First Nations residents of western Canada.

The study examined utilization of acetaminophen with codeine and benzodiazepine drugs among Non-Insured Health Benefits (NIHB) program registered First Nations residents in the four western provinces of Canada and sought preliminary indicators of factors influencing utilization. A small percentage of NIHB clients in the four western provinces were excessive users of acetaminophen with codeine and/or benzodiazepines, but overall utilization of these central nervous system drugs was moderate and within the bounds of non-First Nations population use examined in this study. The study also demonstrated that utilization of acetaminophen with codeine among NIHB claimants was inversely related to the stringency of provincial regulation.

Acetaminophen↗

SALP16, a gene induced in Ixodes scapularis salivary glands during tick feeding.

Guinea pigs infested with Ixodes scapularis acquire antibody-mediated resistance to tick bites, a phenomenon known as tick-immunity. An I. scapularis salivary gland cDNA expression library was therefore probed with sera from tick-immune guinea pigs to identify antigens that elicit humoral responses in the host. Sera from sensitized guinea pigs strongly recognized 3 of 4,500 library clones in an initial screening. The open reading frames of all 3 clones encoded a putative 16.4-kD acidic protein, designated Salp16, with an N-terminal signal sequence and signal peptidase cleavage sites specific for secretory proteins. The salp16 mRNA and Salp16 protein were detected in the salivary glands of engorged, but not unfed, nymphal and adult ticks, and Salp16 was also found in the saliva of engorged ticks. Immunization with recombinant Salp16 induced high antibody titers in guinea pigs, but did not elicit tick-immunity. Salp16 is the first feeding inducible gene that has been cloned from L. scapularis. Molecular characterization of I. scapularis salivary antigens that are induced upon tick feeding should help to facilitate our understanding of tick-host interactions.

Amino Acid Sequence↗

Isolation of West Nile virus from mosquitoes, crows, and a Cooper's hawk in Connecticut.

West Nile (WN) virus, a mosquito-transmitted virus native to Africa, Asia, and Europe, was isolated from two species of mosquitoes, Culex pipiens and Aedes vexans, and from brain tissues of 28 American crows, Corvus brachyrhynchos, and one Cooper's hawk, Accipiter cooperii, in Connecticut. A portion of the genome of virus isolates from four different hosts was sequenced and analyzed by comparative phylogenetic analysis. Our isolates from Connecticut were similar to one another and most closely related to two WN isolates from Romania (2.8 and 3.6 percent difference). If established in North America, WN virus will likely have severe effects on human health and on the health of populations of birds.

Aedes↗