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PubMed2007

Online handwritten shape recognition using segmental hidden Markov models.

We investigate a new approach for online handwritten shape recognition. Interesting features of this approach include learning without manual tuning, learning from very few training samples, incremental learning of characters, and adaptation to the user-specific needs. The proposed system can deal with two-dimensional graphical shapes such as Latin and Asian characters, command gestures, symbols, small drawings, and geometric shapes. It can be used as a building block for a series of recognition tasks with many applications.

Algorithms↗
PubMedDate not supplied

'Do not attempt resuscitation' decision-making: a study exploring the attitudes and experiences of nurses.

At the time of the research being undertaken, a policy regarding resuscitation decisions did not exist at a local hospital. However, it was proposed that a new 'do not attempt resuscitation' policy should be implemented. Nurses' attitudes to, and experiences of, the communication involved in 'do not attempt resuscitation' decision-making were explored and compared with five variables (nurses' age, nurses' grade, years of nursing, area of nursing practice and length of nursing experience on current ward). Seventy-eight qualified nurses, who made up a convenience sample, participated in the study. An adapted questionnaire was used to explore nurses' attitudes to, and experiences of, 'do not attempt resuscitation' decision-making. The findings suggested that nurses' attitudes did generally concur with the guidelines outlined in the new policy. However, in practice, there were many disparities between nurses' experiences of current 'do not attempt resuscitation' decision-making and the policy's guidelines. There were no significant differences between nurses' attitudes to current 'do not attempt resuscitation' decision-making and the five variables. However, there was one significant difference identified between nurses' experiences and the area of nursing practice (p=0.008). To adhere to the principles of the forthcoming 'do not attempt resuscitation' policy at the local hospital, the research findings have suggested that changes need to occur to both nurses' attitudes to and nurses' experiences of current 'do not attempt resuscitation' decision-making.

Adult↗
PubMed2005

The Tayside children's sleep questionnaire: a simple tool to evaluate sleep problems in young children.

BACKGROUND: Behavioural sleep problems in young children are relatively common with between 20% and 40% of those aged 1 to 5 years being affected. This paper describes the development of a simple questionnaire to assess disorders of initiating and maintaining sleep (DIMS) in children aged between 1 and 5 years for use as a selection tool for research purposes or as a screening instrument in primary care. METHODS: A subsection of the Sleep Disturbance Scale for Children was adapted and piloted with a small sample of children in two inner city GP practices (n = 81). Face and content validity were initially established by expert review. Discriminant validity was assessed qualitatively using interviews with mothers of identified cases and non-cases. The validity of the cut-off score was assessed by blinded case note reviews off known cases; inter-rater reliability was also calculated. Following modifications, the final questionnaire was posted to a representative sample of parents across the region with children in the appropriate age band (n = 1023). Internal reliability was assessed using Cronbach's alpha and factor analysis was undertaken to identify significant factors within the questionnaire. RESULTS: The response rate to the population questionnaire was 61.5% (n = 628) with 218 of the children having sleep scores that were indicative of DIMS (35%), echoing other figures reported in the literature. There was good internal consistency for the items (Cronbach's alpha = 0.85) with two main factors accounting for 58% of the variance. CONCLUSION: The Tayside Children's Sleep Questionnaire (TCSQ) is an easy-to-read and reliable tool that could be used both as a clinical and research instrument to assess the severity and prevalence of DIMS in young children.

Child, Preschool↗
PubMed1976

Automated readout of the BG-8 blood-grouping machine. Results of 10,000 blood group and 4,000 rhesus D factor determinations.

An automated readout device recording light transmission in the eight channels of the BG-8, blood-grouping machine after removal of agglutinates is described. Using a continuous recording of positive or negative signals in the eight channels independently, 7.5% of 10,000 blood group determinations had to be retested because of clots in the samples, lacking reactions or excess reactions on the recorder. No incorrect recordings of blood groups were obtained, however. Using anti-D serum together with anti-CDE serum, the rhesus D factor was correctly detected in 99.92% of 3,891 samples, and the Du factor was detected in all 5 samples known to carry this antigen. Using a discontinuous recording of integrated and synchronised signals, adapted to fully automated data processing, another 5% of the samples had to be retested because of inadequate phasing of the reactions in the eight channels of the BQ-8. The results described are influenced by the poor quality of the blood samples used for the study.

ABO Blood-Group System↗
PubMed1991

Iron availability from an infant formula supplemented with bovine lactoferrin.

Iron balance studies were performed in 16 term infants from their 3rd until their 17th week of life. The balance studies were performed at home and comprised five periods with an interval of 3 to 4 weeks, each consisting of three 24-hour collections of milk and stool samples. Seven infants were fed an adapted infant formula supplemented with bovine lactoferrin (100 mg/100 ml) and nine received the same formula without lactoferrin. The lactoferrin supplemented group received 169 micrograms iron/kg b.w. x day and retained 63 micrograms/kg b.w. x day. The mean iron intake of infants fed with the adapted formula without supplementation of lactoferrin was 118 micrograms/kg b.w. x day. The retention of iron was 43 micrograms/kg b.w. x day. Mean percentage retention of iron in the supplemented group was 36%, in the non-supplemented group 28%.

Biological Availability↗
PubMed1978

Simple sensitive microbioassay for adenine arabinoside and hypoxanthine arabinoside in human plasma.

Previous methods using low- or high-pressure liquid chromatography and UV absorbance for quantitation of arabinosides in plasma can practically detect only >/=200 ng of adenine arabinoside and >/=100 ng of hypoxanthine arabinoside per ml, and they require expensive equipment and expert technical assistance. We describe in this report a simple quantitative microbioassay for arabinosides in human plasma based on their ability to inhibit the cytopathic effect of vaccinia virus in an adenosine deaminase-free cell culture system. Using prior separation of nucleosides in plasma by thin-layer chromatography, followed by the microbioassay, we quantitated adenine arabinoside with a sensitivity of approximately 4 ng/ml and hypoxanthine arabinoside at approximately 625 ng/ml. This assay method is simple, sensitive, and reproducible, requires small plasma samples, measures biological activity, and is adaptable to routine use. It is an important tool for evaluating the pharmacology of adenine arabinoside and Ara-AMP in patients in current clinical trials.

Humans↗
PubMed1987

A Combined Immunofluorescence-DNA-Fluorescence Staining Technique for Enumeration of Thiobacillus ferrooxidans in a Population of Acidophilic Bacteria.

An antiserum raised against whole cells of Thiobacillus ferrooxidans was allowed to react with a variety of acidophilic and nonacidophilic bacteria in an enzyme-linked immunosorbent assay and an indirect immunofluorescence assay. Both experiments demonstrated that the antiserum was specific at the species level. This preparation was used to evaluate the role of T. ferrooxidans in the microbial desulfurization process. Leaching experiments were performed, and the numbers of T. ferrooxidans cells and other bacteria were estimated by using a combined immunofluorescence-DNA-fluorescence staining technique that was adapted for this purpose. Nonsterile coal samples inoculated with T. ferrooxidans yielded high concentrations of soluble iron after 16 days. After this period, however, T. ferrooxidans cells could no longer be detected by the immunofluorescence assay, whereas the DNA-fluorescence staining procedure demonstrated a large number of microorganisms on the coal particles. These results indicate that T. ferrooxidans is removed by competition with different acidophilic microorganisms that were originally present on the coal.

Journal Article↗
PubMed2004

Recombination in the genome of Chlamydia trachomatis involving the polymorphic membrane protein C gene relative to ompA and evidence for horizontal gene transfer.

Genome sequencing of Chlamydia trachomatis serovar D has identified polymorphic membrane proteins (Pmp) that are a newly recognized protein family unique to the Chlamydiaceae family. Cumulative data suggest that these diverse proteins are expressed on the cell surface and might be immunologically important. We performed phylogenetic analyses and statistical modeling with 18 reference serovars and 1 genovariant of C. trachomatis to examine the evolutionary characteristics and comparative genetics of PmpC and pmpC, the gene that encodes this protein. We also examined 12 recently isolated ocular and urogenital clinical samples, since reference serovars are laboratory adapted and may not represent strains that are presently responsible for human disease. Phylogenetic reconstructions revealed a clear distinction for disease groups, corresponding to levels of tissue specificity and virulence of the organism. Further, the most prevalent serovars, E, F, and Da, formed a distinct clade. According to the results of comparative genetic analyses, these three genital serovars contained two putative insertion sequence (IS)-like elements with 10- and 15-bp direct repeats, respectively, while all other genital serovars contained one IS-like element. Ocular trachoma serovars also contained both insertions. Previously, no IS-like elements have been identified for Chlamydiaceae. Surprisingly, 7 (58%) of 12 clinical isolates revealed pmpC sequences that were identical to the sequences of other serovars, providing clear evidence for a high rate of whole-gene recombination. Recombination and the differential presence of IS-like elements among distinct disease and prevalence groups may contribute to genome plasticity, which may lead to adaptive changes in tissue tropism and pathogenesis over the course of the organism's evolution.

Bacterial Outer Membrane Proteins↗
PubMed2001

Quantification of human immunodeficiency virus type 1 proviral load by a TaqMan real-time PCR assay.

Proviral human immunodeficiency virus type 1 (HIV-1) DNA could be a useful marker for exploring viral reservoirs and monitoring antiretroviral treatment, particularly when HIV-1 RNA is undetectable in plasma. A new technique was developed to quantify proviral HIV-1 using a TaqMan real-time PCR assay. One copy of proviral HIV-1 DNA could be detected with 100% sensitivity for five copies and the assay had a range of 6 log(10). Reproducibility was evaluated in intra- and interassays using independent extractions of the 8E5 cell line harboring the HIV-1 proviral genome (coefficients of variation [CV], 13 and 27%, respectively) and peripheral blood mononuclear cells (PBMC) from a patient with a mean proviral load of 26 copies per 10(6) PBMC (CV, 46 and 56%, respectively). The median PBMC proviral load of 21 patients, measured in a cross-sectional study, was determined to be 215 copies per 10(6) PBMC (range, <10 to 8,381). In a longitudinal study, the proviral load of 15 out of 16 patients with primary infection fell significantly during 1 year of antiretroviral therapy (P = 0.004). In the remaining patient, proviral HIV-1 DNA was detectable but not quantifiable due to a point mutation at the 5' end of the TaqMan probe. No correlation was observed between proviral load and levels of CD4(+) cells or HIV-1 RNA in plasma. TaqMan PCR is sensitive and adaptable to a large series of samples. The full interest of monitoring proviral HIV-1 DNA can now be ascertained by its application to the routine monitoring of patients.

Anti-HIV Agents↗
PubMed1998

Neutralizing antibodies in sera from macaques immunized with attenuated simian immunodeficiency virus.

Infection with attenuated simian immunodeficiency virus (SIV) in rhesus macaques has been shown to raise antibodies capable of neutralizing an animal challenge stock of primary SIVmac251 in CEMx174 cells that correlate with resistance to infection after experimental challenge with this virulent virus (M. S. Wyand, K. H. Manson, M. Garcia-Moll, D. C. Montefiori, and R. C. Desrosiers, J. Virol. 70:3724-3733, 1996). Here we show that these neutralizing antibodies are not detected in human and rhesus peripheral blood mononuclear cells (PBMC). In addition, neutralization of primary SIVmac251 in human and rhesus PBMC was rarely detected with plasma samples from a similar group of animals that had been infected either with SIVmac239Deltanef for 1.5 years or with SIVmac239Delta3 for 3.2 years, although low-level neutralization was detected in CEMx174 cells. Potent neutralization was detected in CEMx174 cells when the latter plasma samples were assessed with laboratory-adapted SIVmac251. In contrast to primary SIVmac251, laboratory-adapted SIVmac251 did not replicate in human and rhesus PBMC despite its ability to utilize CCR5, Bonzo/STRL33, and BOB/gpr15 as coreceptors for virus entry. These results illustrate the importance of virus passage history and the choice of indicator cells for making assessments of neutralizing antibodies to lentiviruses such as SIV. They also demonstrate that primary SIVmac251 is less sensitive to neutralization in human and rhesus PBMC than it is in established cell lines. Results obtained in PBMC did not support a role for neutralizing antibodies as a mechanism of protection in animals immunized with attenuated SIV and challenged with primary SIVmac251.

Animals↗
PubMed1995

Vasoconstrictor peptides and cold intolerance in patients with stable angina pectoris.

BACKGROUND: The exact mechanism that explains the phenomenon of cold intolerance in patients with angina remains controversial. Although the response to the effects of a cold environment has been examined in these patients, their response to cold air inhalation has produced conflicting results. In addition, the possible role of vasoactive peptides in the pathophysiology has not been explored. OBJECTIVES: The aims of this study were to examine the response of patients with stable angina to the effects of cold air inhalation during exercise testing, and to investigate the possible role played by the vasoconstrictor peptides endothelin-1 (ET-1) and angiotensin-II (AT-II) in the pathophysiology. METHODS: In a randomised order, 12 men with stable angina, whose medication had been stopped, underwent two separate symptom limited treadmill exercise tests. At one visit the patients exercised while breathing room air and at the other visit they exercised while breathing cold air from a specially adapted freezer. Serial peripheral venous blood samples were taken for ET-1 and AT-II estimations during each visit. RESULTS: Cold air inhalation resulted in a significant reduction in the mean time to angina (232.7 (20.4) s v 274.1 (26.9) s, P = 0.04) and the mean total exercise time (299.5 (27.0) s v 350.3 (23.9) s, P = 0.008), but no significant change in the time to 1 mm ST depression (223.3 (29.0) s v 241.3 (29.2) s, P = 0.25). There was no significant difference between the rate-pressure products at the onset of angina (P = 0.13) and the time to 1 mm ST depression (P = 0.85), but at peak exercise the rate-pressure product was significantly lower in patients breathing cold air as opposed to room air (P = 0.049). There was an equivalent significant decrease in ET-1 concentrations at peak exercise compared with that at rest at both visits (room air 5.0 (0.7) pmol/l v 4.3 (0.7) pmol/l, P = 0.03; cold air 4.4 (0.6) pmol/l v 3.8 (0.5) pmol/l, P = 0.02). There was a significant increase in AT-II concentrations 10 min after peak exercise in patients breathing room air (39.2 (6.1) pmol/l v 32.1 (4.8) pmol/l, P = 0.01) which was not repeated during cold air inhalation (36.6 (3.4) pmol/l v 28.3 (3.4) pmol/l, P = 0.07). CONCLUSIONS: Cold air inhalation in patients with stable angina results in an earlier onset of angina and a reduction in exercise capacity. Both peripheral and central reflex mechanisms appear to contribute to the phenomenon of cold intolerance. Peripheral ET-1 and AT-II do not appear to play a significant role in the pathophysiology.

Aged↗
PubMed2005

Prevalence of Fasciola hepatica in dairy herds in England and Wales measured with an ELISA applied to bulk-tank milk.

An ELISA developed to diagnose Fasciola hepatica infection in cattle by detecting serum antibodies was adapted and validated for use with samples of bulk-tank milk. The prevalence of the infection in 61 dairy herds was established by using serum antibody levels or faecal egg counts measured in a proportion of the cows in each herd. The correlation between the results of the ELISA and the herd seroprevalence was 0.83. Using a cut-off value of 27 per cent positive, the bulk-tank ELISA identified herds in which more than 25 per cent of the cows were infected with a diagnostic sensitivity of 96 per cent (95 per cent confidence interval 89 to 100 per cent) and a diagnostic specificity of 80 per cent (95 per cent confidence interval 66 to 94 per cent). By applying the ELISA to 623 herds in England and 445 herds in Wales, the prevalence of F hepatica infection in England was estimated to be 48 per cent (95 per cent confidence interval 46 to 54 per cent), and in Wales 86 per cent (95 per cent confidence interval 84 to 90 per cent).

Animals↗
PubMed1995

Measurement of urinary desmosine as an indicator of acute pulmonary disease.

A modified radioimmunoassay (RIA) for desmosine in the urine was investigated as a tool for the rapid estimation of lung elastin catabolism. Cystic fibrosis (CF) and oxygen toxicity were chosen as conditions that might show altered elastin destruction. Using an antibody bound to magnetic particles the RIA was adapted to handle large numbers of samples requiring only 50 microliters or urine. The experiments show that it was not necessary to hydrolyze or extract the urine prior to assay and that collecting spot urines and normalizing the data to urine creatinine gives the same interpretation of the data as total desmosine in a 24-hour collection. Urine desmosine levels were elevated in 10 of 16 patients with CF; however, daily fluctuations were considerable in some subjects, varying as much as 5-fold and underlining the importance of assaying several consecutive days of urine in acute disorders for an accurate estimate of desmosine excretion. The RIA for desmosine is a rapid and sensitive assay that requires no sample preparation and could be applied to clinical situations that require large numbers of samples.

Acute Disease↗
PubMed1986

The life satisfaction index--well-being: its internal reliability and factorial composition.

The internal consistency and factorial composition of the eight-item Life Satisfaction Index--Well-being, adapted for use with elderly British samples, were examined. Cross-validation was carried out with a randomly drawn community sample of 155 rural people aged 65-89 years. The scale was found to have acceptable internal reliability, but may require modification.

Aged↗
PubMed2005

Adherence with hypertension care among Hmong Americans.

To assess contexts of adherence with hypertension care among Hmong Americans, in-person interviews were conducted with a convenience sample of 323 adults using culturally adapted survey instruments. The mean age of participants was 58 years; 91% had no education, and 86% spoke no English. Although more than 90% had health insurance and were treated with medications, the rate of blood pressure control was low (27%). A majority (> 90%) suffered from psychological distress, and 46% lived with physical illness. Over 50% reported nonadherence with hypertension care. Respondents who were 50 years of age or older, had no physical illness, did not know that hypertension was preventable, or believed that American medicine was too strong, were more likely to report nonadherence with proper medication consumption. Findings suggest that adherence was not due to lack of health care coverage; instead, it may be due to gaps in health services.

Adult↗
PubMed2000

Enzymatic assay of histamine by amperometric detection of H2O2 with a peroxidase-based sensor.

A method for an enzymatic assay of histamine by using histamine oxidase from Arthrobacter globiformis in combination with amperometric determination of H2O2 is described. Histamine could be quantified at a level as low as 10(-7) M. The assay is adaptable to determine histamine in food samples including tuna fish with good sensitivity and selectivity.

Amine Oxidase (Copper-Containing)↗
PubMed1992

Indicators of hospital closure under PPS and Blue Cross/Blue Shield cost-based reimbursements.

Using an adaptation of discriminant analysis on a sample of 120 hospitals in the state of Alabama, this paper illuminates characteristics of hospitals closed due to financial failure under PPS and cost-based Blue Cross/Blue Shield reimbursement systems. The results show high debt levels, poor revenue and cost ratios, poor utilization management, and the inability to collect effectively on outstanding accounts in the year prior to hospital failure.

Alabama↗
PubMed2005

Keratinocyte transient transfections.

The goal of this chapter is to give the reader a concise and easy-to-follow guide to proven transient transfection techniques for primary strains and continuous lines of keratinocytes. The emphasis is on readily available and inexpensive resources that also allow for repeatability and adaptability to scale, experimental conditions, and sample replicates. In addition, basic cell culture techniques necessary to achieve optimal transfection results are provided. Although specified for keratinocytes, these techniques also work well for simple epithelial lines and common laboratory lines, such as COS-7 and CV-1. The standard precipitation protocol is suitable for many experimental designs, and the poly-L-ornithine protocol is intended for situations where calcium is contraindicated. The extended notes section provides necessary technical details to guide the novice user and reminders to the experienced scientist that success with the protocol will depend on several quantitative and qualitative factors including promoter strength, cell culture practices and plasmid isolation.

Calcium Phosphates↗