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Biomedical subjects

K Shaw

Publications and source records attributed to K Shaw.

At least 55 records · Page 3Linked to original sources
PubMed1999

Web site design and development issues: the Washington State Breast and Cervical Health Program Web Site Demonstration Project.

PURPOSE/OBJECTIVES: To explore the development of a customized Web site to assist Breast and Cervical Health Program (BCHP) outreach staff in a community screening program and to evaluate the Internet knowledge and access issues and barriers of outreach staff during a two-year period using the Web site. DESIGN: Knowledge, access issues and barriers, and descriptive questionnaires. SETTINGS: Comprehensive cancer center in Seattle, WA, workshops, and presentations around the state. SAMPLE: BCHP outreach workers, screening coordinators, and almost exclusively public health nurses from regional health districts and program-contracted clinics. METHOD: Web site development was based on continuous input from sample. Detailed descriptions of computer and Internet resources and opinions about the use and usefulness of the BCHP Web site came from a 1996 evaluation and 1998 follow-up conducted using mailed and online Web questionnaires. "Hits" to the Web site were monitored monthly. MAIN RESEARCH VARIABLES: Computer and Internet resources were used along with monthly Web site traffic and opinions about the use and usefulness of the BCHP Web site in the outreach program. FINDINGS: Use of the BCHP Web site has risen steadily over two years to reach a stable plateau. User evaluations show a marked increase in the adoption of the Internet as a working tool. Users believe the Internet is becoming increasingly important to their work. More training and familiarization with the Web is needed. CONCLUSIONS: The Web is an efficient medium for improving communication and providing easy access to resources within the BCHP program. IMPLICATIONS FOR NURSING PRACTICE: Public health programs with meager resources can benefit from the relatively inexpensive use of customized and versatile Web sites.

Breast Neoplasms↗
PubMed1998

Sequence elements involved in the rescue of IBV defective RNA CD-91.

Deletion mutagenesis has been used to identify essential regions for rescue of coronavirus defective RNAs (D-RNAs). Using this technique on a cloned IBV D-RNA CD-91, we have identified a region potentially important in its rescue. Comparing the sequence of D-RNAs rescued with those not rescued we have deduced that a 72 base region corresponding to base number 13,824 to 13,896 in the viral genome is required for rescue. This may be an IBV D-RNA packaging signal or a cis-acting element involved in replication. Further experiments and modification of our techniques will be required to differentiate between the two processes.

Animals↗
PubMed1998

Rescue of IBV D-RNA by heterologous helper virus strains.

Coronavirus defective RNA (D-RNA) vectors could be developed to deliver selected genes for the production of recombinant coronavirus vaccines. An IBV D-RNA, CD-61, derived from a naturally occurring IBV Beaudette D-RNA, CD-91, is being developed as a D-RNA vector for IBV. In order to use CD-61 as a vector it will require rescue by heterologous strains in addition to Beaudette. Rescue will be determined by recognition of replication and packaging signals within the D-RNA by the helper virus. The 5' and 3' UTRs are believed to contain sequences involved in replication and transcription. The 5' and 3' UTRs of six strains of IBV have been sequenced and experiments performed using six strains of helper virus for rescue of CD-61 to determine whether rescue correlates with sequence conservation within the 5' and 3' UTRs. Results indicate that all strains of helper virus rescued the D-RNA to varying degrees. Sequence comparisons show a high degree of sequence identity in the UTRs, but enough strain differences exist to be used as markers. The 5' and 3' UTRs of the D-RNAs rescued by the heterologous strains were also sequenced and leader switching between the helper virus and the Beaudette leader on the D-RNAs was observed.

3' Untranslated Regions↗
PubMed1998

Utilising a defective IBV RNA for heterologous gene expression with potential prophylactic application.

Based on the natural ability of coronaviruses to undergo homologous RNA recombination, we are working to produce infectious bronchitis virus (IBV) recombinants using RNA generated from recombinant fowlpox viruses (FPV). The aim is to replace the spike (S) gene of an existing IBV vaccine strain with the S gene of a heterologous strain. CD-61 is an IBV defective RNA (D-RNA) derived from a naturally occurring IBV D-RNA (CD-91). CD-61 D-RNA is being investigated as an RNA vector for the expression of heterologous genes. T7-derived RNA transcripts of CD-61 can be replicated and passaged in the presence of helper virus, following electroporation into IBV-infected cells. CD-61 cDNA was modified by the addition of the hepatitis delta virus ribozyme plus T7 terminator downstream of the 3'UTR. This allowed the synthesis of discreet RNA transcripts. The complete cassette was cloned into an FPV transfer vector (pEFL10) for generating recombinant fowlpox viruses. FPV/CD-61 recombinants will be assessed for D-RNA production in IBV-infected cells. The luciferase reporter gene sequence has been inserted into the modified CD-61, under the control of the IBV transcription associated sequence (TAS) from gene 5. Luciferase has been successfully expressed from CD-61 in helper virus-infected cells.

Animals↗
PubMed1998

Does IBV change slowly despite the capacity of the spike protein to vary greatly?

We have sequenced that part of the spike protein (S) gene which encodes the aminoterminal and most variable quarter (hypervariable region, HVR) of the S1 subunit of 28 isolates of the 793/B (also known as CR88 and 4/91) serotype of infectious bronchitis virus (IBV) and the whole of S1 for nine of them. The isolates were from France and Britain between the years 1985 (first isolation) and 1996. The maximum nucleotide and amino acid differences between the first isolate and the others were 4.1% and 7.6%, respectively, for the whole of S1 and 7.1% and 14.6%, respectively, in the HVR. Analysis within clearly recognisable subgroups suggested that even in the HVR the nucleotide mutation rate was only 0.3 to 0.6% per year. However, there was no evidence that mutations had become fixed in a progressive manner; this serotype did not appear to be evolving. Strains isolated several years apart could be more similar than those isolated in a given year. It is likely that the amino acid changes are largely at positions where amino acid differences are tolerated rather than as a consequence of immune pressure. Reasons for this conclusion are discussed.

Animals↗
PubMed1998

Fowlpox virus encodes nonessential homologs of cellular alpha-SNAP, PC-1, and an orphan human homolog of a secreted nematode protein.

The genome of fowlpox virus (FWPV), type species of the Avipoxviridae, is considerably rearranged compared with that of vaccinia virus (the prototypic poxvirus and type species of the Orthopoxviridae) and is 30% larger. It is likely that the genome of FWPV contains genes in addition to those found in vaccinia virus, probably involved with its replication and survival in the chicken. A 7,470-bp segment of the FWPV genome has five open reading frames (ORFs), two of which encode ankyrin repeat proteins, many examples of which have been found in poxviruses. The remaining ORFs encode homologs of cellular genes not reported in any other virus. ORF-2 encodes a homolog of the yeast Sec17p and mammalian SNAP proteins, crucial to vesicular transport in the exocytic pathway. ORF-3 encodes a homolog of an orphan human protein, R31240_2, encoded on 19p13.2. ORF-3 is also homologous to three proteins (YLS2, YMV6, and C07B5.5) from the free-living nematode Caenorhabditis elegans and to a 43-kDa antigen from the parasitic nematode Trichinella spiralis. ORF-5 encodes a homolog of the mammalian plasma cell antigen PC-1, a type II glycoprotein with exophosphodiesterase activity. The ORFs are present in the virulent precursor, HP1, of the sequenced attenuated virus (FP9) and are conserved in other strains of FWPV. They were shown, by deletion mutagenesis, to be nonessential to virus replication in tissue culture. RNA encoding the viral homolog of PC-1 is expressed strongly early and late in infection, but RNAs encoding the homologs of SNAP and R31240_2 are expressed weakly and late.

Amino Acid Sequence↗
PubMed1998

Effects of simulated gastroesophageal reflux on the untraumatized rabbit larynx.

OBJECTIVE: Gastroesophageal reflux (GER) has been shown, clinically and experimentally, to cause inflammation of traumatized laryngeal mucosa. This study was performed to determine if GER causes inflammation of untraumatized laryngeal mucosa. METHOD: Sixteen adult New Zealand white rabbits underwent tube pharyngostomy under general anaesthesia without endotracheal intubation. After 7 days recovery, 1 mL/kg of normal saline or HCl (pH 1.5) with pepsin (0.3 mg/mL) was infused into the piriform sinus while the rabbit was under mild sedation: once, twice, or three times per day, for 14 consecutive days. RESULTS: Rabbits that received HCl and pepsin exhibited various degrees of apnea and coughing. Eventually, most developed biphasic stridor on exertion. Histologically, the degree of laryngeal inflammation was greatest when GER was simulated three times per day. Glottic inflammation was greatest in one rabbit, which responded to GER simulation with paroxysmal coughing. CONCLUSION: The mechanism of injury in GER laryngitis is postulated to be a combination of chemical trauma (due to HCl and pepsin) and mechanical trauma (due to an individually variable laryngeal chemoreflex). Further study of GER laryngitis in this new animal model is warranted.

Animals↗
PubMed1997

Transgenic dissection of HIV genes involved in lymphoid depletion.

Transgenic mice carrying an HIV provirus, with selective deletion of all three structural genes, developed extensive lymphoid depletion which was detected not only in the spleen and lymph nodes but also in the thymus. Mice with a high level of HIV gene expression developed acute disease which resulted in premature death, and mice with a low level of viral transcripts developed chronic disease with long-term survival. Neither HIV replication nor the envelope glycoprotein (gp120) was required for T cell depletion. Despite abundant viral gene expression early in life, cell death did not become evident until about the time of full lymphoid maturation, suggesting that thymopoiesis was not affected. The more mature T cells in the peripheral lymphoid organs and in the thymic medulla were less sensitive to the apoptotic process than the immature T cells in the thymic cortex. Gradual depletion of the T cell compartment in the peripheral lymphoid organs was intimately accompanied by the reciprocal expansion of the B cell compartment, resulting in the almost complete replacement of T lymphocytes with B immunoblasts in lymph nodes. Unlike T cells, which showed abundant HIV gene expression, B cells did not. The transgenic approach may help identify the HIV nonstructural gene(s) responsible for immune deficiency and help facilitate dissection of its role in inducing apoptosis.

Animals↗
PubMed1997

Do anomalies of handedness exist in children with Fragile-X syndrome?

Hand preference and skill were assessed in 27 children with Fragile-X syndrome, 30 children with Down's Syndrome, and 29 normal developing children. Three separate and specific anomalies of handedness that have been reported in other learning-disordered groups were examined: increased incidence of left-handedness; ambiguous or inconsistent handedness; and preference-skill discordance. The results revealed no difference between the subject groups in the distribution of left-handedness or ambiguous handedness. The main differences was the percentage of children who were discordant for hand preference and hand skill. In the Fragile-X group 37% preferred to use their right hand but demonstrated no overall difference in skilled performance between the hands. In contrast, only 13% of children with Down's Syndrome and 17% of normal controls showed this pattern of hand discordance. The results are discussed in the context of past research that has shown a similar dissociation of preference and skill in children with autism.

Journal Article↗
PubMed1997

Practice parameters for the assessment and treatment of children and adolescents with anxiety disorders. American Academy of Child and Adolescent Psychiatry.

Anxiety disorders comprise one of the most prevalent categories of psychopathology in children and adolescents. These revised practice parameters highlight the DSM-IV changes for anxiety disorders and review the literature related to the assessment and treatment of anxiety disorders in children and adolescents. Up-to-date information on longitudinal outcome data, assessment of anxiety, parent-child interventions, and use of selective serotonin reuptake inhibitors has been added to the previous parameters, published in September 1993. Recommendations for evaluation and multimodal approaches to treatment are presented.

Anxiety Disorders↗
PubMed1997

The phenomenology of traumatic reactions to psychotic illness.

This study investigated whether a psychotic illness was associated with posttraumatic stress disorder (PTSD) symptomatology in 45 subjects recovering from hospitalization for a psychotic episode. Previous studies have suggested that the experience of psychosis and hospitalization is distressing and that PTSD may be a useful paradigm for the psychological response. Subjects were given questionnaires to evaluate PTSD symptoms, anxiety symptoms, and distress and intrusive memories associated with aspects of treatment and psychosis. Treatment, especially experiences involving a loss of control such as detention, and psychotic symptoms, particularly persecutory delusions, passivity phenomena, and visual hallucinations, were perceived as highly distressing. Twenty-two subjects (52%) met the criteria for a postpsychotic PTSD, with implications for recognition and management of secondary morbidity related to psychosis.

Adaptation, Psychological↗
PubMed1997

II. Localization and characterization of dopamine D4 binding sites in rat and human brain by use of the novel, D4 receptor-selective ligand [3H]NGD 94-1.

The dopamine D4 selective ligand, [H]NGD 94-1, was used in these studies to characterize binding sites in rat and human brain tissue by membrane binding and autoradiography techniques. Autoradiographic analysis of rat brain showed that specific [3H]NGD 94-1 binding was greatest in entorhinal cortex, lateral septal nucleus, hippocampus and the medial preoptic area of the hypothalamus. This nonstriatal distribution of [3H]NGD 94-1 binding was distinct from the autoradiographic distribution of dopamine D2 and D3 receptor subtypes. In homogenate preparations from rat brain regions, [3H]NGD 94-1 binding sites were low in density (<30.0 fmol/mg protein). The low density of D4 binding sites was corroborated by autoradiographic comparisons in which binding density for D4 receptors as measured by [3H]NGD 94-1 was only 1/7 of D2 and 1/5 of D3 receptor densities, despite corrections for differing radioligand binding characteristics. Pharmacological evaluation showed high affinity at rat [3H]NGD 94-1 binding sites for compounds with known D4 receptor affinity and little displacement by compounds with affinity for dopamine D1/D2/D3 receptor subtypes. Specific, high-affinity [3H]NGD 94-1 binding was also present in several human brain regions, including hippocampus, hypothalamus, dorsal medial thalamus, entorhinal cortex, prefrontal cortex and lateral septal nucleus. High-affinity [3H]NGD 94-1 binding was not present in any human striatal region examined. The pharmacological profile of [3H]NGD 94-1 binding sites in human brain was consistent with that previously demonstrated for cloned human D4 receptors expressed in mammalian cells. These findings suggest that specific, high-affinity [3H]NGD 94-1 binding exists in rat and human brain and that these sites reflect populations of dopamine D4 receptors with a distribution unique among dopamine receptor subtypes.

Animals↗
PubMed1997

Towards the routine application of nucleic acid technology for avian disease diagnosis.

The use of nucleic acid technology (polymerase chain reaction, probing, restriction fragment analysis and nucleotide sequencing) in the study of avian diseases has largely been confined to fundamental analysis and retrospective studies. More recently these approaches have been applied to diagnosis and what one might call real-time epidemiological studies on chickens and turkeys. At the heart of these approaches is the identification and characterisation of pathogens based on their genetic material, RNA or DNA. Among the objectives has been the detection of pathogens quickly combined with the simultaneous identification of serotype, subtype or genotype. Nucleic acid sequencing also gives a degree of characterisation unmatched by other approaches. In this paper we describe the use of nucleic acid technology for the diagnosis and epidemiology of infectious bronchitis virus, turkey rhinotracheitis virus (avian pneumovirus) and Newcastle disease virus.

Animals↗
PubMed1996

Identification of pesticide poisoning in wildlife.

The Wildlife Incident Investigation Scheme investigates incidents of suspected poisoning of wildlife (also honey bees and companion animals) by pesticides in the United Kingdom. The approach to these investigations has evolved over the past 30 years. Field investigations, post-mortem examinations, toxicological data and experience of previous poisoning incidents assist in the selection and interpretation of appropriate chemical analyses. Several 'multi-residue' and several 'individual compound' analytical methods for pesticides in wildlife are currently in use; these are described.

Animals↗
PubMed1996

Sequence of the nucleocapsid protein gene of subgroup A and B avian pneumoviruses.

The nucleocapsid protein (N) gene of two subgroup A and one subgroup B strains of avian pneumovirus has been cloned and sequenced. The gene of all three isolates comprised 1197 nucleotides (nt), which formed a single major open reading frame, potentially encoding a protein of 391 amino acid residues. The N gene of the two subgroup A isolates differed by only 1 nt but differed by 282 (24%) nt and 35 (11%) amino acids from the B isolate. The predicted protein was identical in length to that of human, bovine and ovine respiratory syncytial viruses, the amino acid identity being approximately 41% overall but with some regions of identity > 90%.

Amino Acid Sequence↗
PubMed1995

The influence of brain acetaldehyde on oxidative status, dopamine metabolism and visual discrimination task.

The toxic effect of acetaldehyde on brain oxidative capacity and dopamine metabolism has been investigated in rat brains after a single intraperitoneal injection of acetaldehyde (5 mmol/kg) and the results compared with those from chronically ethanol fed rats. Acetaldehyde was present in rat brain 120 hr after a single dose of acetaldehyde, confirming that it is able to cross the blood-brain barrier. Brain catalase increased significantly after acetaldehyde or chronic ethanol administration although there were no other significant changes in the total brain activity of superoxide dismutase, glutathione peroxidase or glutathione reductase. Dopamine turnover was increased in both experimental groups. The acute dose of acetaldehyde reduced the ability of the rats to relearn a computer visual discrimination task.

Acetaldehyde↗
PubMed1995

Generation of a defective RNA of avian coronavirus infectious bronchitis virus (IBV). Defective RNA of coronavirus IBV.

The Beaudette strain of IBV was passaged 16 times in chick kidney (CK) cells. Total cellular RNA was analyzed by Northern hybridization and was probed with 32P-labeled cDNA probes corresponding to the first 2 kb of the 5' end of the genome, but excluding the leader, and to the last 1.8 kb of the 3' end of the genome. A new, defective IBV RNA species (CD-91) was detected at passage six. The defective RNA, present in total cell extract RNA and in oligo-(dT)30-selected RNA from passage 15, was amplified by the reverse transcription-polymerase chain reaction (RT-PCR) to give four fragments. The oligonucleotides used were selected such that CD-91 RNA, but not the genomic RNA, would be amplified. Cloning and sequencing of the PCR products showed that CD-91 comprises 9.1 kb and has three regions of the genome. It contains 1133 nucleotides from the 5' end of the genome, 6322 from gene 1b corresponding to position 12423 to 18744 in the IBV genome and 1626 from the 3' end of the genome. At position 749 one nucleotide, an adenine residue, was absent from CD-91 RNA. By Northern hybridization CD-91 RNA was detected in virions in higher amounts than the subgenomic mRNAs.

Animals↗