Transient Myocardial Thickening in Cats Associated with Heart Failure.

BACKGROUND: Cats with hypertrophic cardiomyopathy (HCM) and congestive heart failure (CHF) can have resolution of both left ventricular hypertrophy and CHF. OBJECTIVES: To describe the clinical characteristics of cats with transient myocardial thickening (TMT) and CHF compared with a control population of cats without resolution of HCM. ANIMALS: A total of 21 cats with TMT, 21 cats with HCM. METHODS: Retrospective study. Clinical records at 4 veterinary centers were searched for TMT cases and a control group of cats with HCM and CHF. TMT was defined as initial maximal left ventricular wall thickness (LVWT) ≥6 mm with left-sided CHF, with subsequent resolution of CHF, reduction in left atrium/aorta (LA/Ao), and LVWT<5.5 mm. HCM was defined as persistent LVWT ≥6 mm. RESULTS: Cats with TMT were younger (2 [0.4-11.4] years) than cats with HCM (8 [1.6-14] years) (P < 0.0001), and antecedent events were more common (15/21 versus 6/21, respectively) (P = 0.01). In cats with TMT, LVWT normalized from 6.8 [6.0-9.7] mm to 4.8 [2.8-5.3] mm and LA/Ao decreased from 1.8 [1.6-2.3] to 1.45 [1.2-1.7] after a mean interval of 3.3 (95% CI: 1.8-4.7) months. CHF recurred in 1 of 21 TMT and 15 of 21 cats with HCM. Cardiac treatment was discontinued in 20 of 21 cats with TMT and 0 of 21 HCM cats. All cats with TMT survived, whereas 8 of 19 cats with HCM died during the study period. CONCLUSIONS AND CLINICAL IMPORTANCE: TMT occurs in younger cats, and antecedent events are common. The prognosis is better in cats with CHF associated with TMT than HCM.

A longitudinal study of 0.2% chlorhexidine gel for removal of mutans streptococci infection in preschool children.

BACKGROUND: The purpose of the present longitudinal study was to determine the effectiveness of 0.2% chlorhexidine (CHX) in removing mutans streptococci (MS) in a treatment cohort of caries-free, preschool children. METHODS: Thirty-six children with mean age 38.6 +/- 15.9 months who had MS infection brushed once daily with 0.2% chlorhexidine (CHX) gel for a period of 6-12 months. The children were examined every three months to assess their oral hygiene and dietary habits and MS status using a commercial microbiological kit. RESULTS: The percentages of children who eliminated MS from their mouths increased from 28 per cent after three months to 48 per cent after six months and over 70 per cent after 12 months of gel use (p = 0.039). Compliance of brushing with CHX gel was significantly associated with MS removal after six months gel use (p < 0.02). At the three and six month recalls, sugar snacks and increased sugar frequency were associated with continued presence of MS (p = 0.03 and p = 0.007). After three months cessation of CHX use, 13 (45 per cent) remained MS free (p = 0.003). CONCLUSIONS: This pilot study involving caries-free preschoolers with MS shows the potential of chlorhexidine gel to remove MS for the limited duration of study.

Factors influencing oral colonization of mutans streptococci in young children.

This paper aims to critically review current knowledge about the key factors involved in oral colonization of the cariogenic group of bacteria, mutans streptococci (MS) in young children. MS, consisting mainly of the species Streptococcus mutans and Streptococcus sobrinus, are commonly cultured from the mouths of infants, with prevalence of infection ranging from around 30 per cent in 3 month old predentate children to over 80 per cent in 24 month old children with primary teeth. MS is usually transmitted to children through their mothers, and the risk of transmission increases with high maternal salivary levels of MS and frequent inoculation. Factors that affect the colonization of MS may be divided into bacterial virulence, host-related and environmental factors. Complex interaction among these factors determine the success and timing of MS colonization in the child. As clinical studies have shown that caries risk is correlated with age at which initial MS colonization occurred, strategies for the prevention of dental caries should include timely control of colonization of the cariogenic bacteria in the mouths of young children.

In vitro and in vivo activities of oligodeoxynucleotide-based thrombin inhibitors containing neutral formacetal linkages.

A series of 15-mer oligodeoxynucleotide analogues were synthesized, and their thrombin inhibitory activities in vitro and in vivo were evaluated. These oligodeoxynucleotide analogues share the same sequence (GGTTGGTGTGGTTGG) but have one or more phosphodiester linkages replaced by a neutral formacetal group. The results obtained from monosubstitutions show that no single phosphodiester group is critical for the thrombin inhibitory activity, suggesting that the interaction between the oligodeoxynucleotide and thrombin is based on a multiple-site charge-charge interaction. Analysis of the effects of different phosphodiester replacements indicates that the backside and left side of the chairlike structure formed by the molecule may be involved in binding with thrombin, presumably by having direct contacts with the anion-binding exosite of the enzyme. For the oligodeoxynucleotides containing two noncontiguous formacetal groups, the effect of the disubstitution is the sum of the effects obtained from the corresponding two monosubstitutions. Infusion of an oligodeoxynucleotide containing four formacetal groups into monkeys showed an increased in vivo anticoagulant effect and an extended in vivo half-life compared to the unmodified oligodeoxynucleotide.

N2- and C8-substituted oligodeoxynucleotides with enhanced thrombin inhibitory activity in vitro and in vivo.

2'-Deoxyguanosine (G) analogues carrying various hydrophobic substituents in the N2 and C8 positions were synthesized and introduced through solid-phase synthesis into 15-mer oligodeoxynucleotide, GGTTGGTGTGGTTGG, which forms a chairlike structure consisting of two G-tetrads and is a potent thrombin inhibitor. The effects of the substitutions at N2 and C8 of the G-tetrad-forming G residues on the thrombin inhibitory activity are relatively small, suggesting that these substitutions cause relatively small perturbations on the chairlike structure formed by the oligodeoxynucleotide. Introduction of a benzyl group into N2 of G6 and G11 and naphthylmethyl groups into N2 of G6 increased the thrombin inhibitory activity, whereas other substituents in these positions had almost no effect or decreased the activity. Particularly, the oligodeoxynucleotide carrying a 1-naphthylmethyl group in the N2 position of G6 showed an increase in activity by about 60% both in vitro and in vivo. Substitutions on the N2 position of other G residues had little effect or decreased the activity. Introduction of a relatively small group, such as methyl and propynyl, into the C8 positions of G1, G5, G10, and G14 increased the activity, presumably due to the stabilization of a chairlike structure, whereas introduction of a large substituent group, phenylethynyl, decreased the activity, probably due to the steric hindrance.

Effects of streptozotocin-induced diabetes on lymphocyte POMC and growth hormone gene expression in the rat.

Diabetes in the rat is associated with a change in the profiles of several neuroendocrine hormones resulting in poor growth and decreased immune function. Since lymphocytes can also serve as a source of neuroendocrine hormones, we have examined whether the change in hormone profiles are accompanied by an impairment of lymphocyte GH and POMC gene expression in the immune system. Diabetes was induced by the administration of streptozotocin (STZ; 10 mg/100 g body weight) and 3 days later GH and ACTH protein and mRNA were determined. The results show a modest diminution of GH RNA in the spleen of diabetic animals whereas the expression of POMC mRNA and ACTH by the thymus was enhanced. The expression of POMC in the spleen appeared unaltered while the increase of POMC RNA in the thymus was evident after the first day of STZ treatment. STZ had no direct effect on GH or POMC expression in the spleen or thymus cells in vitro. Insulin does not appear to be involved in the expression of lymphocyte GH or POMC. The administration of insulin to the diabetic animals had no significant effect on the expression of GH or POMC by the immune cells. In addition, lymphocytes do not appear to serve as a source of insulin or are the expression of genes for lymphocyte GH or ACTH altered by insulin in vitro. Taken together, the findings are the first to report on the expression of neuroendocrine genes in lymphocytes during diabetes. The mechanism for the inhibition of GH and stimulation of POMC expression by lymphocytes in diabetic animals is unknown, but it is tempting to speculate an important role in the development of the autoimmunity that characterizes this complex disease.

Nucleotide-evoked calcium signals and anion secretion in equine cultured epithelia that express apical P2Y2 receptors and pyrimidine nucleotide receptors.

1. Experiments with a spontaneously transformed equine epithelial cell line showed that certain nucleotides increased intracellular free calcium ([Ca2+]i) in cells plated on glass coverslips. The rank order of potency was ATP UTP > 5-Br-UTP, whilst UDP and ADP were ineffective. The response thus appears to be mediated by P2Y2 receptors. 2. Nucleotides also increased short circuit current (Isc) in cells grown into epithelial monolayers and the rank order of potency was UDP> UTP > 5-Br-UTP > ATP > ADP. The increase in [Ca2+]i and the rise in ISC thus have different pharmacological properties. Cross-desensitization experiments indicated that, as well as P2Y2 receptors, the monolayer cultures express at least one additional receptor population that allowed nucleotides to increase ISC. 3. The UDP-evoked increase in ISC was essentially abolished in BAPTA-loaded epithelia suggesting that this response is dependent upon increased [Ca2+]i. Moreover, experiments in which ISC and [Ca2+]i were measured simultaneously showed that the UDP- and ADP-evoked increases in ISC were accompanied by increases in [Ca2+]i. 4. When grown under conditions which favour the development of a polarized phenotype, these epithelial cells thus appear to express [Ca2+]i-mobilizing receptors sensitive to UDP and ADP that are not present in non-polarized cells on coverslips.

Pilot study of the efficacy of a thrombin inhibitor for use during cardiopulmonary bypass.

Heparin is normally used for anticoagulation during cardiopulmonary bypass (CPB), but its use is contraindicated in patients with a history of heparin-induced thrombocytopenia, heparin-provoked thrombosis, or both. Heparin therapy can also be ineffective due to heparin resistance. A short-acting, oligonucleotide-based thrombin inhibitor (thrombin aptamer) may potentially serve as a substitute for heparin in these and other clinical situations. We tested a novel thrombin aptamer in a canine CPB pilot study to determine its anticoagulant efficacy, the resultant changes in coagulation variables, and the aptamer's clearance mechanisms and pharmacokinetics. Seven dogs were studied initially: Four received varied doses of the aptamer (to establish the pharmacokinetic profile) and 3 received heparin. Subsequently, 4 other dogs underwent CPB, receiving a constant infusion of the aptamer before CPB (to characterize the baseline coagulation status), with partial CPB and hemodilution, during 60 minutes of total CPB, and, finally, after a 2-hour recovery period. At a 0.5 mg.kg-1.min-1 dose, the activated clotting time rose with aptamer infusion from 106 +/- 12 seconds to 187 +/- 8 seconds (+/- 1 standard deviation) (p = 0.014), increased further with hemodilution (to 259 +/- 41 seconds; p = 0.017), and was even more prolonged during total CPB (> 1,500 seconds; p < 0.001). This later increase in the activated clotting time paralleled a rise in the plasma concentration of the thrombin aptamer during total CPB, as determined by high-performance liquid chromatography.(ABSTRACT TRUNCATED AT 250 WORDS)

Metaphrase: an aid to the clinical conceptualization and formalization of patient problems in healthcare enterprises.

Patient descriptors, or "problems," such as "brain metastases of melanoma" are an effective way for caregivers to describe patients. But most problems, e.g., "cubital tunnel syndrome" or "ulnar nerve compression," found in problem lists in an Electronic Medical Record (EMR) are not comparable computationally--in general, a computer cannot determine whether they describe the same or a related problem, or whether the user would have preferred "ulnar nerve compression syndrome." Metaphrase is a scalable, middleware component designed to be accessed from problem-manager applications in EMR systems. In response to caregivers' informal descriptors it suggests potentially equivalent, authoritative, and more formally comparable descriptors. Metaphrase contains a clinical subset of the 1997 UMLS Metathesaurus and some 10,000 "problems" from the Mayo Clinic and Harvard Beth Israel Hospital. Word and term completion, spelling correction, and semantic navigation, all combine to ease the burden of problem conceptualization, entry and formalization.

Cupped lesions of early onset dental erosion in young southeast Queensland adults.

BACKGROUND: Dental erosion manifests as cupped lesions on cusp apices and in fissures of teeth in patients from southeast Queensland referred with excessive tooth wear. When found in young adults, these lesions may indicate early onset of active dental erosion. If the numbers and extent of cupped lesions increase with age, erosion may be a slow cumulative process. METHODS: This cross-sectional study recorded the presence or absence and the relative sizes of cupped lesions from all cusps and occlusal fissures on premolar and permanent molar teeth from study models by image analysis. Type-specimens of cupped lesions were examined. RESULTS: The incidence by tooth reflected time in the mouth, post-tooth emergence. A linear increase in lesion number and size, with age, was found. However, cupped lesions occurred on mandibular first molar cusp apices as often, and attained greater extent, in adults under 27 years compared with older subjects. CONCLUSION: Marked differences were found between lesion number and size, between maxillary and mandibular molar sites that reflect differences in salivary protection against dental erosion. The significance of this study is that the mandibular first permanent molar indicates the age of onset and severity of dental erosion.

An evaluation of UMLS as a controlled terminology for the Problem List Toolkit.

We are developing a set of software components--the Problem List Toolkit (PL-Tk)--to support operations on clinical problem labels. An adaptation of the National Library of Medicine's Unified Medical Language System (UMLS) provides general vocabulary services to domain-specific software components. Our initial investigation centers on the inclusion in UMLS of problem labels used in the Beth Israel Deaconess Medical Center's Online Medical Record (OMR). We also explore the semantic typing of problem labels matched in UMLS. We have operationally defined a clinical problem to derive its semantic type from classes of terms representing findings or processes typically requiring diagnostic evaluation or therapeutic management in clinical practice. Of 1262 unique OMR problem labels, 999 terms (79%) have matches in UMLS. 986 of 999 terms (99%) map to the UMLS concept of the corresponding lexical match. 952 of 999 terms (95%) have semantic types that comply with our operational definition of clinical problems. These 952 terms (75%) constitute Version 1.0 of the problem list vocabulary B196. Matching terms with inappropriate semantic types raise issues regarding requirements for PL-Tk, typing of existing UMLS terms, and the adequacy of our operational definition for clinical problems. UMLS provides a large repertoire of pre-coordinated terms that are used as problem labels in a heavily used computer-based patient record system. The semantic type hierarchy provides a framework for the consistent use of clinical concepts in problem lists such that clinical problem labels represent "good" clinical problems.

Gender differences in the sensitivity to negative stimuli: cross-modal affective priming study.

BACKGROUND: There is evidence showing that men and women differ with regard to the processing of emotional information. However, the mechanisms behind these differences are not fully understood. METHOD: The sample comprised of 275 (167 female) right-handed, healthy participants, recruited from the community. We employed a customized affective priming task, which consisted of three subtests, differing in the modality of the prime (face, written word, and sound). The targets were always written words of either positive or negative valence. The priming effect was measured as reaction time facilitation in conditions where both prime and target were emotional (of the same positive or negative valence) compared with conditions where the emotional targets were preceded by neutral primes. RESULTS: The priming effect was observed across all three modalities, with an interaction of gender by valence: the priming effect in the emotionally negative condition in male participants was stronger compared with females. This was accounted for by the differential priming effect within the female group where priming was significantly smaller in the emotionally negative conditions compared with the positive conditions. The male participants revealed a comparable priming effect across both the emotionally negative and positive conditions. CONCLUSION: Reduced priming in negative conditions in women may reflect interference processes due to greater sensitivity to negative valence of stimuli. This in turn could underlie the gender-related differences in susceptibility to emotional disorders.

Handheld Flyback driven coaxial dielectric barrier discharge: Development and characterization.

The development of a handheld single and triple chamber atmospheric pressure coaxial dielectric barrier discharge driven by Flyback circuitry for helium and argon discharges is described. The Flyback uses external metal-oxide-semiconductor field-effect transistor power switching technology and the transformer operates in the continuous current mode to convert a continuous dc power of 10-33 W to generate a 1.2-1.6 kV 3.5 micros pulse. An argon discharge breakdown voltage of approximately 768 V is measured. With a 50 kHz, pulse repetition rate and an argon flow rate of 0.5-10 argon slm (slm denotes standard liters per minute), the electrical power density deposited in the volume discharge increases linearly at a rate of 75+/-20% mW/cm(3) per 1 slm of gas. Electrical power transfer efficiency between the secondary Flyback coil and the discharge volume increases from 0.1% to 0.65%. Neutral argon gas forced convection analysis yields a similar energy loss rate to the electrical discharge process. Optical emission spectroscopy studies of the expanding discharge plume into ambient air reveal that the air climatically controls the plume chemistry to produce an abundance of neutral argon atoms and molecular nitrogen.

A component-based problem list subsystem for the HOLON testbed. Health Object Library Online.

One of the deliverables of the HOLON (Health Object Library Online) project is the specification of a reference architecture for clinical information systems that facilitates the development of a variety of discrete, reusable software components. One of the challenges facing the HOLON consortium is determining what kinds of components can be made available in a library for developers of clinical information systems. To further explore the use of component architectures in the development of reusable clinical subsystems, we have incorporated ongoing work in the development of enterprise terminology services into a Problem List subsystem for the HOLON testbed. We have successfully implemented a set of components using CORBA (Common Object Request Broker Architecture) and Java distributed object technologies that provide a functional problem list application and UMLS-based "Problem Picker." Through this development, we have overcome a variety of obstacles characteristic of rapidly emerging technologies, and have identified architectural issues necessary to scale these components for use and reuse within an enterprise clinical information system.

Validation of clinical problems using a UMLS-based semantic parser.

The capture and symbolization of data from the clinical problem list facilitates the creation of high-fidelity patient resumes for use in aggregate analysis and decision support. We report on the development of a UMLS-based semantic parser and present a preliminary evaluation of the parser in the recognition and validation of disease-related clinical problems. We randomly sampled 20% of the 26,858 unique non-dictionary clinical problems entered into OMR (Online Medical Record) between 1989 and August, 1997, and eliminated a series of qualified problem labels, e.g., history-of, to obtain a dataset of 4122 problem labels. Within this dataset, the authors identified 2810 labels (68.2%) as referring to a broad range of disease-related processes. The parser correctly recognized and validated 1398 of the 2810 disease-related labels (49.8 +/- 1.9%) and correctly excluded 1220 of 1312 non-disease-related labels (93.0 +/- 1.4%). 812 of the 1181 match failures (68.8%) were caused by terms either absent from UMLS or modifiers not accepted by the parser; 369 match failures (31.2%) were caused by labels having patterns not recognized by the parser. By enriching the UMLS lexicon with terms commonly found in provider-entered labels, it appears that performance of the parser can be significantly enhanced over a few subsequent iterations. This initial evaluation provides a foundation from which to make principled additions to the UMLS lexicon locally for use in symbolizing clinical data; further research is necessary to determine applicability to other health care settings.

The simultaneous measurement of epithelial ion transport and intracellular free Ca2+ in cultured equine sweat gland secretory epithelium.

We explored the relationship between nucleotide-evoked changes in intracellular free calcium ([Ca2+]i) and anion secretion by measuring [Ca2+]i and I(SC) simultaneously in Fura-2-loaded, cultured equine sweat gland epithelia. Apical ATP, UTP or UDP elicited sustained increases in [Ca2+]i that were initiated by the mobilization of cytoplasmic Ca2+ but maintained by Ca2+ influx. However, although these nucleotides also increased I(SC), this response was transient whereas the [Ca2+]i signals were sustained. Experiments in which external Ca2+ was removed/replaced showed that Ca2+ entering nucleotide-stimulated cells elicited very little change in ISC. Cross desensitization experiments showed that UTP-stimulated epithelia became insensitive to ATP but that UTP could increase both [Ca2+]i and ISC in ATP-stimulated cells by activating 'pyrimidinoceptors' essentially insensitive to ATP. Thapsigargin evoked a sustained rise in [Ca2+]i that was accompanied by a maintained increase in ISC. However, this increase in ISC was dependent upon external Ca2+ and so the responses to nucleotides and thapsigargin have different properties. ATP increased ISC in thapsigargin-treated cells without causing any rise in [Ca2+]i while ionomycin increased both parameters. The data therefore show that apical P2Y receptors allow nucleotides to increase ISC via two mechanisms, one of which appears to be [Ca2+]i-independent control of anion channels.

A peptide derived from a tissue factor loop region functions as a tissue factor--factor VIIa antagonist.

Tissue factor (TF) is a transmembrane protein that functions in the initiation of blood coagulation in vivo. At sites of vascular injury, TF serves as a cell-surface receptor for the serine protease factor VIIa (FVIIa), forming an enzyme--cofactor complex and enhancing the catalytic activity of FVIIa. Tissue factor, along with the receptors for alpha- and gamma-interferons, is a member of the class 2 cytokine receptor superfamily. Crystallographic analysis demonstrated that the extracellular domain of TF consists of two immunoglobulin-like domains joined by a linker region. Each domain is comprised of two antiparallel beta-sheets containing seven conserved beta-strands separated by more variable loop regions. Extensive mutagenesis has been performed in order to map the FVIIa binding site on TF. Results indicated that the discontinuous binding site for FVIIa lies at the domain--domain interface and includes residues from extended loops and beta-strands within both the N- and C-terminal domains. Our previous study provided evidence that three consecutive residues (D44, W45, K46) within the TF loop region between beta-strands C and C' of the N-terminal domain were important for interactions with FVIIa. We have presently extended our alanine-scanning mutagenesis to include the residues within the flanking beta-strands. Thirteen sTF mutants were screened for their ability to enhance FVIIa activity. Three residues within strand C (Y34, Q37, I38) and two residues within C' (K48, Y51) were shown to be important for TF cofactor function.(ABSTRACT TRUNCATED AT 250 WORDS)

Stimulation of chloride secretion by baicalein in isolated rat distal colon.

The effect of baicalein on mucosal ion transport in the rat distal colon was investigated in Ussing chambers. Mucosal addition of baicalein (1-100 microM) elicited a concentration-dependent short-circuit current (I(sc)) response. The increase in I(sc) was mainly due to Cl(-) secretion. The presence of mucosal indomethacin (10 microM) significantly reduced both the basal and subsequent baicalein-evoked I(sc) responses. The baicalein-induced I(sc) were inhibited by mucosal application of diphenylamine-2-carboxylic acid (100 microM) and glibenclamide (500 microM) and basolateral application of chromanol 293B (30 microM), a blocker of K(v)LQT1 channels and Ba(2+) ions (5 mM). Treatment of the colonic mucosa with baicalein elicited a threefold increase in cAMP production. Pretreating the colonic mucosa with carbachol (100 microM, serosal) but not thapsigargin (1 microM, both sides) abolished the baicalein-induced I(sc). Addition of baicalein subsequent to forskolin induced a further increase in I(sc). These results indicate that the baicalein evoked Cl(-) secretion across rat colonic mucosa, possibly via a cAMP-dependent pathway. However, the action of baicalein cannot be solely explained by its cAMP-elevating effect. Baicalein may stimulate Cl(-) secretion via a cAMP-independent pathway or have a direct effect on cystic fibrosis transmembrane conductance regulator.

In vitro uptake of bromodeoxyuridine by human nasopharyngeal carcinoma (NPC) and its relation to clinical findings.

A cell kinetic study of 27 newly diagnosed patients with nasopharyngeal carcinoma (NPC) using the in vitro bromodeoxyuridine (BrdU) technique was performed. The results were reproducible as demonstrated by three independent sections performed on each patient. No correlation between BrdU labelling index (LI) and Ho's clinical staging was found. A higher LI was associated with the development of distant metastases (P = 0.057). Statistically significant correlation was found between low LI and longer duration required to achieve complete remission in the primary site of disease (P = 0.026). This study suggests a potential role for in vitro BrdU labelling index as a prognosticator for NPC prior to treatment.

Conversion of thrombin into an anticoagulant by protein engineering.

At sites of vascular injury, thrombin interacts with multiple procoagulant substrates, to mediate both fibrin clotting and platelet aggregation. But upon binding to thrombomodulin on the vascular endothelium, thrombin instead activates protein C, thereby functioning as an anticoagulant and attenuating clot formation. Upon infusion in vivo, both the procoagulant and anticoagulant effects of thrombin were observed. Preliminary studies indicating that thrombin's protein C activating and fibrinogen clotting activities could be dissociated by mutagenesis suggested to us that a thrombin variant that lacked procoagulant activity while retaining anticoagulant function might be an attractive antithrombotic agent. Using protein engineering, we introduced a single substitution, E229A, that substantially shifted thrombin's specificity in favour of the anticoagulant substrate, protein C. In monkeys, this modified thrombin functioned as an endogenous protein C activator demonstrating dose-dependent, reversible anticoagulation without any indication of procoagulant activity. Notably, template bleeding times were not prolonged, suggesting a reduced potential for bleeding complications.