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Biomedical subjects

W Woods

Publications and source records attributed to W Woods.

At least 19 recordsLinked to original sources

Recombinant NY-ESO-1 cancer antigen: production and purification under cGMP conditions.

The cancer-testis antigen, NY-ESO-1, has been engineered into a bacterial expression plasmid which incorporates a His6-tag. The plasmid was transfected into E. coli strain BL21 and Master and Working cell banks generated from this expression system. Three 15-litre fermentations were performed under cGMP (code of Good Manufacturing Practice) conditions and the crude NY-ESO-1 tagged protein isolated as solubilised inclusion bodies. A three-step cGMP chromatography process (immobilised metal affinity, anion exchange, and hydrophobic interaction) was utilised to purify the protein. The purified NY-ESO-1 is being used in early stage human cancer vaccine trials in Australia and the U.S.A.

Antigens, Neoplasm↗

Neuroblastomas of infancy exhibit a characteristic ganglioside pattern.

BACKGROUND: Gangliosides are membrane-bound glycolipid molecules particularly prominent in neural tissue. Changes in ganglioside expression during embryologic development result from a shift in biosynthesis from the fetal b pathway to the adult a pathway. Tumor gangliosides may play a role in the clinical behavior of certain subtypes of neuroblastoma. Because neuroblastoma, which presents in infancy, has a different biologic and clinical phenotype than that which presents in older children, the authors determined whether differences in ganglioside biosynthesis exist between these two neuroblastoma subgroups. METHODS: Sixty-eight tumor specimens (25 diagnosed by screening and 43 diagnosed clinically) were obtained from the Quebec Neuroblastoma Screening Project. Gangliosides were isolated and purified by solvent partitioning, separated by high performance thin-layer chromatography, and quantitated by scanning densitometry. The sum of a and b pathway gangliosides were determined for each tumor. RESULTS: Gangliosides of the b (fetal) pathway predominated in both screened and clinically diagnosed tumors of patients younger than 1 year of age. Twenty-three of 25 screened patients (92%) and 21 of 23 patients with clinically diagnosed tumors at younger than 1 year of age (91%) had tumor b pathway ganglioside content greater than 60%. In contrast, tumors of only 8 of 20 patients 1 year or older (40%) had b pathway ganglioside predominance. Predominance of b pathway tumor gangliosides correlated with improved outcome. Event free survival was significantly higher among patients with b pathway ganglioside tumor content greater than 60% versus those with b pathway ganglioside tumor content less than 60% (118.1 +/- 3.9 months vs. 69.2 +/- 8.6 months, P < 0.01). CONCLUSIONS: Fetal patterns of ganglioside biosynthesis predominate in neuroblastoma tumors from patients younger than 1 year of age and adult patterns of ganglioside biosynthesis predominate in tumors from older children, supporting the view that neuroblastoma consists of distinct but overlapping disorders, and that gangliosides may play a biologic role in the clinical differences among these patients.

Chromatography, Thin Layer↗

Plasma pharmacokinetics and cerebrospinal fluid penetration of thioguanine in children with acute lymphoblastic leukemia: a collaborative Pediatric Oncology Branch, NCI, and Children's Cancer Group study.

PURPOSE: In preclinical studies, thioguanine (TG) has been shown to be more potent than the standard acute lymphoblastic leukemia (ALL) maintenance agent, mercaptopurine (MP), suggesting that TG may be more efficacious than MP in the treatment of childhood ALL. As part of a pilot trial in which TG was used in place of MP, we studied the plasma pharmacokinetics of oral TG and measured steady-state plasma and CSF TG concentrations during a continuous intravenous infusion (CIVI) in children with newly diagnosed standard-risk ALL. METHODS: Nine plasma samples were collected after each patient's first 60 mg/m2 oral TG dose during maintenance. CIVI TG (20 mg/m2/h over 24 h) was administered during the consolidation phase of therapy, and simultaneous plasma and CSF samples were collected near the end of the infusion. TG was measured by reverse-phase HPLC with ultraviolet detection. Erythrocyte TG nucleotide (TGN) concentrations were measured 7 days after a course of CIVI TG and prior to the start of each maintenance cycle. RESULTS: After oral TG (n = 35), the mean (+/- SD) peak plasma concentration was 0.46 +/- 0.68 microM and the AUC ranged from 0.18 to 9.5 microM.h (mean 1.5 microM.h). Mean steady-state plasma and CSF TG concentrations during CIVI (n = 33) were 2.7 and 0.5 microM, respectively. The mean (+/- SD) TG clearance was 935 +/- 463 ml/min per m2. Plasma TG concentrations did not correlate with erythrocyte TGN concentrations after oral or CIVI TG. The 8-OH-TG metabolite was detected in plasma and CSF. CONCLUSIONS: TG concentrations that are cytotoxic to human leukemia cell lines can be achieved in plasma after a 60 mg/m2 oral dose of TG and in plasma and CSF during CIVI of TG.

Administration, Oral↗

The Newcastle Auditory Battery (NAB). A temporal and spatial test battery for use on adult naïve subjects.

A battery of tests for assessing the perception of temporal and spatial acoustic cues is described, together with a software platform for implementing the battery. The software runs on a personal computer either with a sound card or with widely used laboratory hardware. The battery is intended for use with neurologically impaired and other naive subjects, to allow inference at the single-subject level for any given subtest. The aim is to allow a systematic psychoacoustic evaluation of complex sound processing in single patients. Normal values are given for the threshold data for 30 naïve control subjects aged from 20 to 60 years. Future modifications of the battery are allowed by modular software architecture.

Adult↗

Oxacillin susceptibility testing of Staphylococcus saprophyticus using disk diffusion, agar dilution, broth microdilution, and the Vitek GPS-105 card.

Eighty-three mecA negative isolates of S. saprophyticus had oxacillin zone diameters <or= 15 mm or MICs ranging from <or= 0.25-1.0 microg/ml when tested by either agar dilution, broth microdilution, or the Vitek GPS-105 card. Greater than 90% of these isolates would be considered resistant using NCCLS M7-A5, M100-S10 criteria. These results suggest that the current NCCLS MIC and zone diameter breakpoints for oxacillin resistance in coagulase-negative Staphylococci are not appropriate for S. saprophyticus as they do not correlate with the presence of the mecA gene.

Bacterial Proteins↗

Comparison of phenotypic methods to identify enterococci intrinsically resistant to vancomycin (VanC VRE).

Clinical laboratories must be able to differentiate between enterococci possessing acquired resistance to vancomycin (vanA and vanB genotypes) from those that are inherently resistant (vanC1 and vanC2/C3 genotypes). We compared several routine phenotypic tests to determine the species identity of clinical isolates of enterococci and a PCR assay for the van ligase genes was used to confirm identification of VanC VRE. The Vitek Gram Positive Identification card identified 53/60 (88%) Enterococcus faecalis and E. faecium isolates and 81/141 (57%) VanC VRE without additional testing. Another 32 of the VanC VRE required additional testing (e.g., motility and pigmentation) for correct identification. However, 7 of these 32 VanC VRE were nonmotile. The rapid ID 32 STREP strips identified 50/60 (83%) E. faecalis and E. faecium isolates and 102/141 (72%) VanC VRE. All E. faecalis and E. faecium isolates were nonmotile and did not acidify 1% methyl-alpha-D-glucopyranoside (MGP). Only 93/115 (81%) E. gallinarum and 21/26 (81%) E. casseliflavus/E. flavescens were motile but all 141 VanC VRE acidified MGP. MGP acidification can accurately differentiate VanC VRE from E. faecalis and E. faecium. Because some VanC VRE isolates are nonmotile, MGP acidification is preferred as a simple and less costly test for identification of these isolates.

Bacterial Proteins↗

Genotyping Taenia tapeworms by single-strand conformation polymorphism of mitochondrial DNA.

To overcome limitations in identifying tapeworms of the genus Taenia by traditional approaches, we have established a single-strand conformation polymorphism (SSCP) method utilizing two different regions of mitochondrial (mt) DNA as targets. The NADH dehydrogenase 1 and the cytochrome c oxidase subunit I genes were amplified from genomic DNA by polymerase chain reaction (PCR), denatured and subjected to electrophoresis in mutation detection enhancement gels. SSCP analysis achieved delineation among eight different species of Taenia from different hosts based on characteristic profiles and enabled the detection of intraspecific variability in profiles for some taxa. This SSCP-based typing method has important implications for taxonomy, diagnosis and for studying the genetic structure of Taenia populations.

Animals↗

Inhibition of PCR in genital and urine specimens submitted for Chlamydia trachomatis testing.

We determined the frequency of PCR inhibition in genital and urine specimens submitted for Chlamydia trachomatis testing using the internal control DNA provided with the COBAS AMPLICOR C. trachomatis test and assessed methods to remove it. Inhibition occurred in 65 of 906 (7%) cervical swabs, 23 of 51 (45%) urethral swabs, and 2 of 175 (1.1%) urine samples. Overall, inhibition was eliminated in processed specimens after storage at 4 degrees C in 77 of 90 specimens (86%), freezing at -70 degrees C in 59 of 82 specimens (72%), storage at 4 degrees C followed by either 1:100 dilution in 37 of 43 specimens (86%) or 1:10 dilution in 42 of 47 specimens (89%), and phenol-chloroform extraction in 79 of 80 specimens (99%). No positive specimens were missed due to inhibition. We conclude that PCR inhibition is rare with urine specimens and infrequent with endocervical swabs but occurs frequently with urethral swabs. The frequency of PCR inhibition may be significantly reduced by methods which can be easily incorporated into the processing of specimens.

Automation↗

Clinical and epidemiological significance of enterococci intrinsically resistant to vancomycin (possessing the vanC genotype)

Constitutive low-level vancomycin resistance is found intrinsically in certain enterococcal species and is encoded by vanC ligase genes. These intrinsically vancomycin-resistant enterococci (VRE) will be referred to as VANC VRE. A prospective study to determine the clinical and epidemiologic significance of VANC VRE was conducted. VANC VRE were recovered from the stools of 34 of 601 (5.7%) patients, a rate similar to that obtained for the stools of 100 outpatients in the community (5%). VANC VRE were also isolated from the nonstool specimens of 9 of 538 patients (1.7%), including two patients with bacteremia. No VRE of the vanA or vanB genotypes were detected in nonstool specimens. Eighty-two hospital contacts of the first 23 patients found to be colonized or infected with VANC VRE were screened, and 6 contacts were found to be gastrointestinal carriers of VANC VRE. However, typing of isolates from these 6 contacts by pulsed-field gel electrophoresis with SmaI showed the isolates to be unique and different from those recovered from the index patients. In fact, all VANC VRE isolates from different patients in this study were unique. A case-control study with patients who were negative when screened for VANC VRE as controls failed to identify any risk factor associated with colonization or infection with this organism. VANC VRE were infrequently recovered from clinical specimens but were occasionally found as part of the normal stool flora. Since no transmission between patients was documented, additional isolation procedures may not be necessary for patients colonized or infected with VANC VRE.

Adult↗

Revised nucleotide sequence of an archaeal insertion element (ISH28) reveals a putative transposase gene.

The published sequence of the insertion element ISH28 contained many small ORFs that were difficult to interpret. We resequenced the entire element and found seven nucleotide differences. The corrected sequence of ISH28 is 938 bp long, and now reveals a single open reading frame of 828 bp. The putative protein is highly similar (49% aa identity) to the predicted transposase of ISH1.

Amino Acid Sequence↗

Probable new species of Desulfovibrio isolated from a pyogenic liver abscess.

A fastidious, slowly growing, spiral gram-negative bacterium was isolated from the liver abscess of an 82-year-old man with a 3-week history of febrile illness. The organism was an obligate anaerobe that grew at 37 and 42 degrees C but not at 25 degrees C. Its vibrioid or spiral morphology on Gram staining, rapid progressive motility, electron micrograph features, and biochemical tests were all consistent with the organism belonging to the genus Desulfovibrio. 16S rRNA gene sequencing of this organism demonstrated a 97% similarity to Desulfovibrio desulfuricans with 45 nucleotide differences, suggesting that it is a new species of Desulfovibrio.

Aged↗

Persistence and possible progression of a pelvic neuroblastoma detected by mass screening during 19 months.

PURPOSE: The preclinical detection of neuroblastoma by screening for elevated levels of urinary catecholamines often leads to the discovery of children with early-stage, biologically favorable disease. It is uncertain how vigorously therapy must be pursued in such cases. We report an infant whose pelvic mass was initially thought to be a fecaloma, and consequently was not treated for 19 months. CASE REPORT: A 2-month-old girl was referred for evaluation for the presence of a neuroblastoma because of elevated urinary catecholamines detected in a mass screening program. Although no mass was initially found, urinary catecholamines became increasingly elevated, and a pelvic mass was finally radiologically identified. It was resected. Histology showed it to be a neuroblastoma, POG stage C, International stage 3, with unfavorable Shimada features, although near triploid with an unamplified N-myc oncogene. Chemotherapy was given for five cycles, and the child remains well, with no evidence of disease. CONCLUSION: Although mass screening programs often detect early- stage, biologically favorable neuroblastomas that may spontaneously regress, our case had rising levels of urinary catecholamines and unfavorable histologic features at the time of resection. The unusual location of the tumor, and radiologic features of a fecaloma, contributed to the delay in definitive diagnosis, although persistence of elevated urinary catecholamines in the absence of a tumor is infrequent.

Catecholamines↗

The jobs Americans hold.

How many Americans hold which jobs? What careers have futures? Grouped here are all the jobs employing more than 150,000 people--they include about 90% of U.S. workers--and the growth that the Bureau of Labor Statistics calculates for each.

Data Collection↗

Donor and host influences in bone marrow transplantation for immunodeficiency disease and leukemia.

Bone marrow transplantation is an effective form of therapy for lethal immunodeficiency diseases and leukemia. Patients who are treated by bone marrow transplantation for these diseases have an improved outcome if treated early after diagnosis, before they have developed secondary complications. Recent advances in transplantation have allowed choices between several donor types. These alternative donor types are the subject of this analysis from the University of Minnesota. In these diseases, matched sibling donor bone marrow transplantation is the standard for comparison. In individuals with lethal immunodeficiencies (severe combined immune deficiency [SCID], Wiskott-Aldrich syndrome [WAS], Chédiak-Higashi syndrome [CHS]) who lack a sibling donor, unrelated transplantation has produced results that are almost equal to those of matched sibling transplants. Patients with high-risk acute lymphoblastic leukemia (ALL) who have received sibling or unrelated transplants have results that are superior to autologous donor transplants. In ALL, there is a need to use new therapies, eg, immunotoxins, to decrease the currently high relapse rate. In patients with acute myelogenous leukemia (AML) in first complete remission, results are excellent and comparable using related and autologous donors. Results in non-first-remission AML are inferior and do not differ if related, unrelated, or autologous transplants are used. In chronic myelogenous leukemia (CML), early survival results are superior using autologous and related transplants as compared with unrelated transplants.

Bone Marrow Transplantation↗