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K Mills

Publications and source records attributed to K Mills.

At least 37 records · Page 2Linked to original sources
PubMed2001

Evaluation of Etest method for determining caspofungin (MK-0991) susceptibilities of 726 clinical isolates of Candida species.

The performance of the Etest for testing the susceptibilities to caspofungin (MK-0991) of 726 isolates of Candida spp. was assessed against the National Committee for Clinical Laboratory Standards (NCCLS) microdilution broth method. The NCCLS method employed RPMI 1640 broth medium, and MICs were read after incubation for 48 h at 35 degrees C. MICs were determined by Etest for all 726 isolates with RPMI agar containing 2% glucose (RPG) and were read after incubation for 48 h at 35 degrees C. The Candida isolates included Candida albicans (n = 486), Candida glabrata (n = 96), Candida tropicalis (n = 51), Candida parapsilosis (n = 47), Candida krusei (n = 11), Candida lusitaniae (n = 2), and Candida guilliermondii (n = 33). In addition, a subset of 314 isolates were also tested by Etest using Casitone agar (CAS) and antibiotic medium 3 agar (AM3). The Etest results obtained using RPG correlated well with reference MICs. Overall agreement was 94% with RPG, 82% with CAS, and 79% with AM3. When RPG was used, agreement ranged from 79% for C. parapsilosis to 100% for C. krusei, C. lusitaniae, and C. guilliermondii. When CAS was used, agreement ranged from 0% for C. lusitaniae to 100% for C. glabrata. With AM3, agreement ranged from 0% for C. lusitaniae to 100% for C. guilliermondii. All three media supported growth of each of the Candida species. Etest results were easy to read, with sharp zones of inhibition. In most instances (75%) where a discrepancy was observed between the Etest and the reference method, the Etest MIC was lower. The Etest method using RPG appears to be useful for determining caspofungin susceptibilities of Candida species.

Anti-Bacterial Agents↗
PubMed2001

Long term outcome in Lambert-Eaton myasthenic syndrome without lung cancer.

OBJECTIVES: To determine the prognosis in patients with Lambert-Eaton myasthenic syndrome (LEMS) without small cell lung cancer (SCLC), and to analyse longitudinal clinical, electrophysiological, and immunological data on each patient to establish prognostic factors for long term outcome. METHODS: The retrospective and part prospective study of 47 patients with LEMS was undertaken from data recorded during visits to a specialist neuromuscular clinic. Serial measurements of muscle strength score in shoulder abduction, elbow extension and hip flexion, compound muscle action potential (CMAP) amplitude, and postcontraction increment in abductor digiti minimi (ADM), and anti-P/Q-type voltage gated calcium channel (VGCC) antibody titre were made at each visit. RESULTS: Muscle strength scores were improved in 88% of patients after a median duration of immunosuppressive treatment of 6 years (range 1.3 to 17 years); anti-VGCC antibody titres fell in 52% after treatment; and mean resting CMAP amplitude improved from 2.7 mV initially to 8.8 mV after 2 years of treatment p<0.001). Initial pretreatment anti-VGCC antibody titre did not correlate significantly with either CMAP amplitude, CMAP increment, or clinical score: from serial measurements made during follow up, significant correlation between antibody titre and CMAP amplitude was seen in only two patients. Sustained clinical remission was achieved by 20 (43%) of whom only four remained in remission without the need for immunosuppression. Using a Cox proportional hazards model, the only independent predictor of sustained clinical remission was initial pretreatment clinical score (p=0.03). Lymphoma presented in three patients during the study. CONCLUSIONS: The prognosis in patients with LEMS without SCLC is favourable, although patients often need significant doses of immunosuppressive treatment to remain clinically stable. Only initial clinical muscle strength measurements and not anti-VGCC antibody titres or electrophysiological recordings are predictive of long term outcome.

Adolescent↗
PubMed2001

Simple, directional cDNA cloning for in situ transcript hybridization screens.

Here, we describe a suppression PCR-based method for directional cloning of randomly primed cDNAs from small quantities of tissue. Synthesis of the first cDNA strand is conducted on oligonucleotide-coated magnetic beads. Synthesis of the second strand is accomplished using nonspecifically primed suppression PCR. This method is used to synthesize a cDNA library from zebrafish embryos at 6-9 h after fertilization. The sequencing of the clones and their use in an in situ hybridization screen to detect restricted patterns of gene transcription in zebrafish embryos showed that this method allows the rapid identification of genes that are important for development and genes that are expressed at levels undetectable by whole-mount in situ transcript hybridization. The random priming of cDNA alleviates the problems encountered in the identification of zebrafish genes from poly(dT)-primed cDNA clones caused by the long 3' UTRs frequently found in transcripts from this organism.

Animals↗
PubMed2001

Identification of alpha(1)-antitrypsin variants in plasma with the use of proteomic technology.

BACKGROUND: Proteomic technology permits the investigation of genetic metabolic diseases at the level of protein expression. Changes in the expression, polypeptide structure, and posttranslational modification of individual proteins can be detected in complex mixtures of proteins. METHODS: We used high-resolution two-dimensional polyacrylamide gel electrophoresis to separate isoforms of plasma proteins and detect abnormalities of mass and/or charge. We confirmed the identity of the separated proteins by in-gel digestion with proteases and N-glycanases and then analyzed the released peptides and glycans by matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry. RESULTS: Complete characterization of the polypeptide sequences and glycosylation of alpha(1)-antitrypsin isoforms was achieved in plasma from controls and from patients with three different known alpha(1)-antitrypsin deficiencies and congenital disorder of glycosylation type Ia. CONCLUSIONS: This study shows that proteomic techniques are a powerful and sensitive means of detecting changes in the amino acid sequence and abnormal posttranslational modifications of specific proteins in a complex biologic matrix.

Amino Acid Sequence↗
PubMed2000

An essential role for the caspase dronc in developmentally programmed cell death in Drosophila.

Dronc is a caspase recruitment domain-containing Drosophila caspase that is expressed in a temporally and spatially restricted fashion during development. Dronc is the only fly caspase known to be regulated by the hormone ecdysone. Here we show that ectopic expression of dronc in the developing fly eye leads to increased cell death and an ablated eye phenotype that can be suppressed by halving the dosage of the genes in the H99 complex (reaper, hid, and grim) and enhanced by mutations in diap1. In contrast to previous reports, we show that the dronc eye ablation phenotype can be suppressed by coexpression of the baculoviral caspase inhibitor p35. Dronc also interacts, both genetically and biochemically, with the CED-4/Apaf-1 fly homolog, Dark. Furthermore, extracts made from Dark homozygous mutant flies have reduced ability to process Dronc, showing that Dark is required for Dronc processing. Finally, using the RNA interference technique, we show that loss of Dronc function in early Drosophila embryos results in a dramatic decrease in cell death, indicating that Dronc is important for programmed cell death during embryogenesis. These results suggest that Dronc is a key caspase mediating programmed cell death in Drosophila.

Animals↗
PubMed2000

The discovery and synthesis of highly potent, A2a receptor agonists.

A series of N6,2-disubstituted adenosine analogues have been synthesized and their functional activity measured against A2a and A1 receptors. Examples of compounds with both a lipophilic N6-substituent and amino-functionalized 2-position were highly active at the A2a receptor on the human neutrophil.

Adenosine↗
PubMed2000

The use of refusal postcards in recruiting older adults.

This article examines whether a refusal postcard makes recruitment more efficient or instead reduces response rates to a telephone survey of older adults. Medicare health maintenance organization (HMO) members were randomly sampled in sequential phases. All samples received an initial contact letter from a HMO geriatrician. A refusal postcard was included in the first sample (N = 178); however, the remaining six samples did not receive this postcard (N = 1,003). An overall refusal rate of 32% was observed when postcards were included versus a 14% rate of refusal when postcards were excluded (p < .001). When potential respondents were reached by telephone, refusal rates were similar (9% versus 10%). Despite the higher refusal rate among the sample receiving the refusal postcard, no significant differences in demographics, health, and health behaviors were observed between the two final sample groups completing the survey. We conclude that refusal postcards greatly increase the refusal rates without offering any prescreening advantage in the recruitment process of older adults and could increase the costs of recruitment for a telephone survey. Furthermore, use of a refusal postcard precludes individuals from making fully informed decisions about participating in research.

Aged↗
PubMed2000

The mitochondrial trifunctional protein: centre of a beta-oxidation metabolon?

The trifunctional enzyme comprises three consecutive steps in the mitochondrial beta-oxidation of long-chain acyl-CoA esters: 2-enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase and 3-ketoacyl-CoA thiolase. Deficiencies in either 3-hydroxyacyl-CoA dehydrogenase activity, or all three activities, are important causes of human disease. The dehydrogenase and thiolase have a requirement for NAD+ and CoA respectively, whose levels are conserved within the mitochondrion and thus provide possible means for control and regulation of beta-oxidation. Using analysis of the intact CoA ester intermediates produced by the complex, we have examined the sensitivity of the complex to NAD+/NADH and acetyl-CoA. We consider the evidence for channelling within the trifunctional protein and propose a model for a beta-oxidation 'metabolon'.

Amino Acid Sequence↗
PubMed2000

Evaluation of the Etest method for determining voriconazole susceptibilities of 312 clinical isolates of Candida species by using three different agar media.

Performance of the Etest for voriconazole susceptibility testing of 312 isolates of Candida spp. was assessed against that of the National Committee for Clinical Laboratory Standards (NCCLS) microdilution broth method. The NCCLS method employed RPMI 1640 broth medium, and MICs were read after incubation for 48 h at 35 degrees C. Etest MICs were determined with RPMI agar containing 2% glucose (RPG), Casitone agar (CAS), and antibiotic medium 3 (AM3) agar and were read after incubation for 48 h at 35 degrees C. The Candida spp. isolates included C. albicans (n = 174), C. glabrata (n = 55), C. tropicalis (n = 31), C. parapsilosis (n = 39), C. krusei (n = 5), C. lusitaniae (n = 2), and C. guilliermondii (n = 6). The Etest results obtained using RPG correlated well with the reference MICs. Overall agreement ranged from 91% for C. glabrata to 100% for C. tropicalis, C. parapsilosis, C. guilliermondii, C. krusei, and C. lusitaniae. When CAS was used, agreement ranged from 80% for C. krusei to 100% for C. parapsilosis, C. guilliermondii, and C. lusitaniae. With AM3, agreement ranged from 58% for C. glabrata to 100% for C. lusitaniae and C. guilliermondii. The Etest method using RPG appears to be a useful method for determining voriconazole susceptibilities of Candida species.

Agar↗
PubMed2000

In vitro susceptibility testing of filamentous fungi: comparison of Etest and reference microdilution methods for determining itraconazole MICs.

The performance of the Etest for itraconazole susceptibility testing of 50 isolates of filamentous fungi was assessed in comparison with the National Committee for Clinical Laboratory Standards (NCCLS) proposed standard microdilution broth method. The NCCLS method employed RPMI 1640 broth medium, and MICs were read after incubation for 48 h at 35 degrees C. Etest MICs were determined with RPMI agar containing 2% glucose and with Casitone agar and were read after incubation for 24 h (Aspergillus spp. and Rhizopus spp.) and 48 h (all species except Rhizopus spp.) at 35 degrees C. The isolates included Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Aspergillus terreus, Fusarium spp., Pseudallescheria boydii, Rhizopus spp., Paecilomyces variotii, and an Acremonium sp. Overall agreement between Etest and microdilution MICs was 96% with RPMI agar and 80% with Casitone agar. The agreement was 100% for all species except Rhizopus spp. (83%) and Paecilomyces varioti (0%) with RPMI agar. When Casitone agar was used, the agreement ranged from 50% with Rhizopus spp. to 100% with Fusarium spp., P. boydii, P. varioti, and an Acremonium sp. Notably, for Aspergillus spp., the agreement between itraconazole Etest MICs read at 24 h and reference microdilution MICs read at 48 h was 100% with both RPMI and Casitone agar. Both media supported the growth of all filamentous fungi tested. Where a discrepancy was observed between Etest and the reference method, the Etest MIC was generally higher. The Etest method using RPMI agar appears to be a useful method for determining itraconazole susceptibilities of Aspergillus spp. and other filamentous fungi.

Antifungal Agents↗
PubMed2000

Association of Moraxella ovis with keratoconjunctivitis in mule deer and moose in Wyoming.

Six cases of infectious keratoconjunctivitis (IKC) in mule deer (Odocoileus hemionus) and moose (Alces alces) in Wyoming (USA) were investigated during fall and winter of 1995 and 1996. Excessive lacrimation, mucopurulent conjunctivitis, keratitis, and corneal opacity were observed in mule deer. Moose had severe mucopurulent conjunctivitis, keratitis, and corneal ulceration. Hemolytic, non-piliated Moraxella ovis was isolated from two mule deer and two moose. We attempted to reproduce IKC in three mule deer fawns using an isolate of M. ovis from a clinically affected mule deer. These fawns did not develop clinical signs of infection and the bacterium was not reisolated from inoculated deer. Inoculated deer may not have developed clinical signs because deer were not exposed to ultraviolet light or mechanical insult before inoculation. In addition, the isolate used for inoculation may have lost virulence factors through passage, or M. ovis may not have been the primary pathogen responsible for clinical disease in the natural cases of IKC we investigated. The etiology of IKC in free-ranging wild ruminants remains poorly understood.

Animals↗
PubMed2000

Bacteria and nematodes in the conjunctiva of mule deer from Wyoming and Utah.

Swabs of conjunctiva were collected from 44 live and 226 hunter-harvested mule deer (Odocoileus hemionus) from Wyoming and Utah (USA). We identified 29 gram negative and 22 gram positive bacterial taxonomic categories, but many isolates from hunter-harvested animals were environmental contaminants. Staphylococcus spp. and Micrococcus spp. were the most common gram positive bacteria isolated, and Enterobacter spp., Escherichia coli, and Pseudomonas spp. were common gram negative bacteria isolated. Thelazia californiensis were found in 15% of hunter-harvested deer in Utah in 1994 and in 8% in 1995. Nematodes were found in 40% of live deer in 1995 and 66% in 1996. Three live animals showed clinical signs of infectious keratoconjunctivitis (IKC) in 1996, but pathogenic bacteria were not isolated from these individuals. Hemolytic, non-piliated Moraxella ovis was isolated from two clinically normal live deer in 1996 and isolates were similar to those cultured from IKC cases from Wyoming and Utah.

Animals↗
PubMed1999

Changes in motor evoked potentials to short-interval paired transcranial magnetic stimuli in multiple sclerosis.

OBJECTIVE: Paired transcranial magnetic stimuli (TMS) were applied in 8 multiple sclerosis (MS) patients with asymmetrical clinical signs and in 8 healthy controls to test the hypothesis that the circuits responsible for the generation and transmission of I-waves are abnormal in the former group METHODS: A figure-of-8 coil discharging through a Magstim 200/Bistim configuration delivered identical stimuli at an intensity 10% above the motor threshold of the relaxed first dorsal interosseous muscle. The interstimulus intervals (ISIs) used were varied in a pseudo-randomized fashion in steps of 0.2 ms between 1.0 and 5 ms. RESULTS: In 9 of 12 unilateral studies in the control group, a pattern of 3 peaks of increased motor evoked potential size was found at ISIs of 1.2-1.6 ms, 2.4-3.2 ms and 4.4-5.0 ms. A similar pattern was present in only 5 of 12 studies in the patients (Fisher's exact test, P = 0.1), while it was absent in all the 4 studies of the side with greater clinical involvement in patients (P = 0.01) CONCLUSION: Our results suggest that I-wave generation is more likely to be defective in MS than in normal subjects, that this defect resides in the cortex, and that it correlates with severity of physical signs.

Adult↗
PubMed1999

T cell proliferation, MHC class II restriction and cytokine products of gliadin-stimulated peripheral blood mononuclear cells (PBMC).

The immune response of PBMC to gliadin was investigated in patients with coeliac disease (CoD) by examining proliferation, MHC restriction and cytokine production. Gliadin induced low levels of proliferation in 63% of eight untreated patients, 32% of 28 treated patients and 35% of 31 healthy control subjects. In MHC restriction studies, the proliferative response to gliadin was inhibited (range 47-98% inhibition) in the presence of a MoAb to HLA-DR in each of three coeliac and three control donors studied. Using flow cytometry, increased expression of activation markers (HLA-DR and IL-2R) was demonstrated on gliadin-stimulated T cells from four of nine coeliac patients and three of seven healthy control donors. Cytokines were studied in culture supernatants using ELISA. Gliadin was a potent inducer of IL-6 and IL-10 in 100% of coeliac patients and controls, whereas IL-4 was not produced in either subject group. Gliadin induced IL-2 production in 40% of untreated patients, 42% of treated patients and 35% of healthy control donors. Interferon-gamma (IFN-gamma) in gliadin-stimulated cultures was found only in coeliac patients, observed in 33% of untreated patients and 25% of treated patients. Spontaneous secretion of both IL-2 and IFN-gamma was found more frequently in patients with untreated disease (87% of cases versus 21% of controls for IFN-gamma and 40% versus 0% for IL-2). These results suggest, as manifest by IFN-gamma production, that gliadin stimulates a Th1/Th0-like response in coeliac patients and a Th0-like response in healthy controls.

Adolescent↗
PubMed1999

Transmission of tuberculosis from a seven-year-old child in a Sydney school.

OBJECTIVES: To determine whether a 7-year-old child with extrapulmonary and pulmonary tuberculosis (TB) and direct smear positive sputum for acid-fast bacilli was infectious to home and school contacts, and to ascertain potential adult sources of infection for these contacts. METHODS: Contact tracing by Mantoux testing was conducted on 220 children at a primary school and after-school care facility, and 59 selected adults considered potential sources of infection. RESULTS: The participation rate for the children was 98% and 92% for the adults. Mantoux positivity (induration >/= 10 mm, or >/= 15 mm with previous BCG) among children was 13% at the school (anticipated rate 2-3%), 26% among school staff, and 7% among children at the after-school care centre where the index case attended. One exposed adult hospital staff member converted from Mantoux negative to positive. No other cases of TB disease were detected among children or adults tested. CONCLUSION: Although spread of TB from children to others is rare, the findings of this investigation indicate that transmission of TB from a young child to other children and an adult may have occurred, and that sputum testing and contact tracing for sputum smear positive children should be considered.

Adult↗
PubMed1999

Molecular analysis of the group 1 and 2 allergens from the house dust mite, Euroglyphus maynei.

BACKGROUND: There is increasing evidence that the house dust mite Euroglyphus maynei may be a significant source of allergic sensitization. The structural information for the E. maynei allergens is largely restricted to a single partial genomic sequence of Eur m 1. METHODS: A cDNA library was constructed from a culture of E. maynei. Clones encoding the major group 1 and 2 allergens were isolated by DNA hybridization and sequenced. RESULTS: The sequence of several full length clones of Eur m 1 and Eur m 2 were obtained. The full pre-proenzyme sequence of the cysteine protease Eur m 1 was determined. The translated amino acid sequence of Eur m 1 and Eur m 2 had 84-86% sequence identity with the corresponding allergens from Dermatophagoides pteronyssinus and Dermatophagoides farinae mites. This is the same as the degree of sequence identity found between D. pteronyssinus and D. farinae despite Euroglyphus being a member of the Pyroglyphinae subfamily rather than the Dermatophagoidinae subfamily. CONCLUSION: The sequences of the major Eur m 1 and Eur m 2 allergens are described. Their degree of divergence from the Dermatophagoides spp. is similar to that observed between D. pteronyssinus and D. farinae group 1 and group 2 allergens.

Allergens↗
PubMed1999

Medications frequently used for asthma and rhinitis. Asthma Steering Committee, Penn State Geisinger Health System.

Multiple medications are available to treat asthma and other allergic diseases, making it difficult to determine which medications to use. The decision should be based on efficacy, cost, and other advantages and disadvantages. Comparative tables can help for rapid selection and can be used as a tool for enhancing prescribing practices. This article organizes this information into a tabular format, allowing for rapid review.

Anti-Asthmatic Agents↗