The use of fluorescence lifetime technology in benign and malignant thyroid tissues.

OBJECTIVE: To explore the use of fluorescence lifetime imaging microscopy in thyroid tissues, and to investigate how different thyroid lesions affect fluorescence lifetime. METHOD: Fluorescence lifetime measurements were taken of fresh frozen thyroid surgical specimens stained with fluorescein isothiocyanate tagged anti-thyroglobulin monoclonal antibodies. RESULTS: The mean fluorescence lifetime measurements in 12 patients - 3 with multinodular goitre, 4 with follicular adenoma, 4 with papillary thyroid carcinoma and 1 with follicular carcinoma - were 3.16 ns (range, 2.66-3.52 ns), 3.75 ns (range, 2.99-4.57 ns), 2.97 ns (range, 2.57-3.21 ns) and 3.61 ns, respectively. The fluorescence lifetime of follicular adenoma patients was higher than that of papillary thyroid carcinoma patients by 26 per cent (p = 0.058). The fluorescence lifetime in the follicular carcinoma patient was similar to the follicular adenoma group, but higher than in the papillary thyroid carcinoma group by 22 per cent (p = 0.01). CONCLUSION: Fluorescence lifetime measurements varied in different thyroid pathologies, possibly because of tissue-scale structural influences.

Cardioprotection from stress conditions by weak magnetic fields in the Schumann Resonance band.

The Schumann Resonances (ScR) are Extremely Low Frequency (ELF) electromagnetic resonances in the Earth-ionosphere cavity excited by global lightning discharges. This natural electromagnetic noise has likely existed on the Earth ever since the Earth had an atmosphere and an ionosphere, hence surrounding us throughout our evolutionary history. The purpose of this study was to examine the influence of extremely weak magnetic fields in the ScR first mode frequency range on the spontaneous contractions, calcium transients and Creatine Kinase (CK) release of rat cardiac cell cultures. We show that applying 7.8 Hz, 90 nT magnetic fields (MF) causes a gradual decrease in the spontaneous calcium transients' amplitude, reaching 28% of the initial amplitude after 40 minutes of MF application, and accompanied with a gradual decrease in the calcium transients' rise time. The mechanical spontaneous contractions cease after the ScR fields have been applied for more than 30 minutes, when the calcium transient's amplitude reached ~60% of its initial value. The influence of the ScR MF was reversible, independent of the field magnitude in the range 20 pT-100 nT, and independent of the external DC magnetic field. However, the effect is frequency dependent; the described changes occurred only in the 7.6-8 Hz range. In addition, applying 7.8 Hz, 90 nT MF for 1.5 hours, reduced the amount of CK released to the buffer, during normal conditions, hypoxic conditions and oxidative stress induced by 80 µM H2O2. We show that the ScR field induced reduction in CK release is associated with a stress response process and has a protective character.

Diffusion Reflection: A Novel Method for Detection of Oral Cancer.

Intraoperative detection of residual disease in oral cancer may reduce the high rate of recurrences. The aim of the present study was to evaluate the detection sensitivity of diffusion reflection (DR) measurements of bioconjugated gold nanorods (GNRs) to cancerous sites in a rat model of oral squamous cell carcinoma. We used hyperspectral spectroscopy and DR measurements of GNRs bioconjugated to slide specimens of rat tongues where squamous carcinoma was induced by 4NQO (4-nitroquinoline-N-oxide). Wistar-derived male rats were used: 6 were sacrificed at wk 32 to 37 following 4NQO administration (experimental rats), as were 2 control rats at wk 32 and 36. The detection results were compared with histopathology: 19 sites of cancerous changes were identified microscopically (11 invasive cancer and 8 carcinoma in situ [CIS]). The GNRs attached selectively to areas of carcinomatous changes with an intensity exceeding 17 intensity units at 780 nm (overall specificity, 97%; overall sensitivity, 87%) when the hyperspectral spectroscopy system was used. The resulting DR slopes of the reflected intensity showed an increase of >80% in areas of invasive cancer and an increase of >30% in the CIS sites. The resulting intensity units of the hyperspectral spectroscopy system in the invasive cancer significantly exceed those of the CIS (t test, p = .0002; Mann-Whitney, p = .0024). The results demonstrate a great potential of the direct DR scanning as a new and simple tool for detecting residual disease intraoperatively.

Relation between ambient air quality and selected birth defects, seven county study, Texas, 1997-2000.

A population-based case-control study investigated the association between maternal exposure to air pollutants, carbon monoxide, nitrogen dioxide, ozone, sulfur dioxide, and particulate matter <10 microm in aerodynamic diameter during weeks 3-8 of pregnancy and the risk of selected cardiac birth defects and oral clefts in livebirths and fetal deaths between 1997 and 2000 in seven Texas counties. Controls were frequency matched to cases on year of birth, vital status, and maternal county of residence at delivery. Stationary monitoring data were used to estimate air pollution exposure. Logistic regression models adjusted for covariates available in the vital record. When the highest quartile of exposure was compared with the lowest, the authors observed positive associations between carbon monoxide and tetralogy of Fallot (odds ratio = 2.04, 95% confidence interval: 1.26, 3.29), particulate matter <10 microm in aerodynamic diameter and isolated atrial septal defects (odds ratio = 2.27, 95% confidence interval: 1.43, 3.60), and sulfur dioxide and isolated ventricular septal defects (odds ratio = 2.16, 95% confidence interval: 1.51, 3.09). There were inverse associations between carbon monoxide and isolated atrial septal defects and between ozone and isolated ventricular septal defects. Evidence that air pollution exposure influences the risk of oral clefts was limited. Suggestive results support a previously reported finding of an association between ozone exposure and pulmonary artery and valve defects.

Differential aspects in ratio measurements of [Ca(2+)](i) relaxation in cardiomyocyte contraction following various drug treatments.

This study is concerned with the analysis of the time dependency of [Ca(2+)](i), monitored by indo-1-AM, via the ratiometric time response curve R(t) as measured during contractions of spontaneous or electrical stimulated cardiomyocytes (in culture). A mathematical formulation which describes the relaxation phase of R(t) was developed. By fitting formulation to the measured data of R(t), the extraction of characteristic parameters is feasible, which may reflect the factors regulating intracellular Ca concentration. The usefulness of the suggested formulation was examined by monitoring changes induced in those parameters following the exposure of the myocytes to different drugs, among which are: caffeine, ryanodine, thapsigargin db, cyclic AMP, isoprenaline, doxorubicin, and Cl-IB-MECA.

Analysis of early apoptotic events in individual cells by fluorescence intensity and polarization measurements.

Apoptosis is a dynamic process of variable duration. The ability to continuously detect the death process occurring in single or subgroups of cells is therefore very important in identifying apoptotic cells within a complex population. The Individual Cell Scanner (ICS), a multiparametric, multilaser-based scanning static cytometer, was used in the present report for the continuous monitoring of the apoptosis process. Fluorescence intensity (FI), polarization (FP), kinetic measurements, and cluster analysis of subpopulations were carried out utilizing various fluorescent probes. Hydrogen peroxide-induced apoptosis was monitored online in intact live lymphocytes by continuous sequential measurements of intracellular hyperpolarization. Plasma membrane asymmetry, mitochondrial membrane potential, and lysosomal rupture were monitored in individual cells. Cytoplasmic condensations, due to cell shrinkage and early lysosomal rupture, were found to be very early events of apoptosis. The new analytical capabilities suggested here may provide simple and convenient methods for detecting apoptosis from its earlier stages.

Effect of center volume on outcome of ventricular septal defect closure and arterial switch operation.

This study evaluates the effects of surgical volume and preoperative patient characteristics on length of stay following ventricular septal defect (VSD) repair and arterial switch operation (ASO). Twenty-four centers contributed data on 16,795 consecutive surgical procedures from January 1, 1992 to December 31, 1996 to the Pediatric Cardiac Care Consortium Registry. The following variables were used in the analysis: center, center volume, age, weight, date of operation, preoperative hospital days, presence of trisomy 21, and significant coexisting heart defects (risk). For VSD, all variables were associated with length of stay in the univariate analyses; however, in the multivariate model, center volume did not retain significance. Length of stay for ASO was associated with center, center volume, and preoperative days in the univariate analysis. In the multivariate analysis for ASO, center and center volume were significant variables for predicting length of stay. These findings suggest that length of stay for uncomplicated surgical procedures is not related to surgical volume; however, for technically demanding procedures, length of stay tends to be shorter in centers with more experience.

The role of the right ventricle during hypobaric hypoxic exercise: insights from patients after the Fontan operation.

OBJECTIVES: The principal objective of this study was to examine the importance of the right ventricle for maximal systemic oxygen transport during exercise at high altitude by studying patients after the Fontan operation. BACKGROUND: High-altitude-induced hypoxia causes a reduction in maximal oxygen uptake. Normal right ventricular pump function may be critical to sustain cardiac output in the face of hypoxic pulmonary vasoconstriction. We hypothesized that patients after the Fontan operation, who lack a functional subpulmonary ventricle, would have a limited exercise capacity at altitude, with an inability to increase cardiac output. METHODS: We measured oxygen uptake (VO2, Douglas bag), cardiac output (Qc, C2H2 rebreathing), heart rate (HR) (ECG), blood pressure (BP) (cuff), and O2 Sat (pulse oximetry) in 11 patients aged 14.5+/-5.2 yr (mean +/- SD) at 4.7+/-1.6 yr after surgery. Data were obtained at rest, at three submaximal steady state workrates, and at peak exercise on a cycle ergometer. All tests were performed at sea level (SL) and at simulated altitude (ALT) of 3048 m (10,000 ft, 522 torr) in a hypobaric chamber. RESULTS: At SL, resting O2 sat was 92.6+/-4%. At ALT, O2 sat decreased to 88.2+/-4.6% (P < 0.05) at rest and decreased further to 80+/-6.3% (P < 0.05) with peak exercise. At SL, VO2 increased from 5.1+/-0.9 mL x kg(-1) x min(-1) at rest to 23.5+/-5.3 mL x kg(-1) x min(-1) at peak exercise and CI (Qc x m(-2)) increased from 3.3+/-0.7 L x m(-2) to 6.2+/-1.2 L x m(-2). VO2 peak, 17.8+/-4 mL x kg(-1) x min(-1) (P < 0.05), and CI peak, 5.0+/-1.5 L x m(-2) (P < 0.05), were both decreased at ALT. Remarkably, the relationship between Qc and VO2 was normal during submaximal exercise at both SL and ALT. However at ALT, stroke volume index (SVI, SV x m(-2)) decreased from 37.7+/-8.6 mL x min(-1) x m2 at rest, to 31.3+/-8.6 mL x min(-1) x m2 at peak exercise (P < 0.05), whereas it did not fall during sea level exercise. CONCLUSIONS: During submaximal exercise at altitude, right ventricular contractile function is not necessary to increase cardiac output appropriately for oxygen uptake. However, normal right ventricular pump function may be necessary to achieve maximal cardiac output during exercise with acute high altitude exposure.

Prenatal exposures and congenital heart defects in Down syndrome infants.

The purpose of this study was to determine whether there are important differences in maternal and environmental prenatal risk factors between liveborn Down syndrome infants with congenital heart defects and Down syndrome infants without heart defects. Using a case control study design, we evaluated the risk associated with maternal illness, drug ingestion, substance usage, and chemical exposures in the home or workplace. The period of risk selected was 3 months before and 3 months after the last menstrual period, because cardiac development occurs early, before the mother may become aware of her pregnancy. Because fetal survival in Down syndrome may be more vulnerable to various exposures, controls were selected who also had trisomy 21. Of 171 infants studied, 89 were cases with congenital heart disease, and 82 were controls without heart disease. All interviews were performed by one nurse practitioner using a structured standardized questionnaire. Cases and controls had similar maternal ages, family incomes, parental education levels, and contraceptive practices before pregnancy. No differences were found between case and control mothers for maternal illness, medication use, or consumption of caffeinated beverages, cigarettes, or alcohol. Reporting of recreational drug usage was infrequent, may reflect underreporting, and did not differ between cases and controls. Maternal exposures were commonly reported for pesticides (50%), hair dyes (22%), craft paints (8%), varnishes (7%), and solvents (3.5%). However, in none of the categories was maternal exposure significantly more prevalent among case mothers than among control mothers. The failure of this study to identify risk factors for cardiac malformations may be attributable to the small differences in reported frequencies reducing statistical power or to the possibility that cardiac malformation in Down syndrome is a direct result of chromosomal duplication.

Prelytic stimulation of target and effector cells following conjugation as measured by intracellular fluorescein fluorescence polarization.

The aim of the present study was to detect prelytic intracellular changes induced in target and effector cells following their conjugation at room temperature. Changes in the cytoplasmic matrix were measured by means of intracellular fluorescein fluorescence polarization (IFFP) using the Cellscan apparatus. Both natural killer and lymphocyte activated killer cells were used as effector cells, while K562 and Daudi cell lines were used as targets. The results show that following their conjugation, both the effector and the target cells show significant reductions (>10%) in IFFP values. Changes in IFFP were induced by specific interaction and only between viable cells. No evidence of fluorescein transfer from a stained cell to its nonstained counterpart was found. To the best of our knowledge, this is the first time that effector-target interaction is monitored on an individual cell basis within a population, by means of IFFP measurements. In addition, in order to explain the physical phenomena, measurements of physical parameters which might affect the IFFP, such as changes in osmolality and pH, were performed and discussed. © 1998 Society of Photo-Optical Instrumentation Engineers.

Influenza A virus affects the response of human peripheral blood mononuclear cells to phytohaemagglutinin A by altering the cytoskeleton.

In the present study, we demonstrate that the infection of human peripheral blood mononuclear cells (PBMC) with influenza A virus caused changes in intracellular fluorescein fluorescence polarization (IFFP) which, as previously described, reflect alterations in the polymerization of the cytoskeleton. Kinetic measurements revealed two cycles of an approximate 10% decrease in IFFP within 3.5 and 5 h after infection. Infection win influenza A virus also altered the response of PBMC to phytohaemagglutinin (PHA), which was manifested as changes of 5.3 and 4% in IFFP at 1 and 2 h after infection, respectively. the changes in IFFP correlated with DNA synthesis measured 72 h after exposure to PHA. These results show the ability of IFFP measurements to identify early intracellular metabolic events induced in virus-infected cells.

Monitoring of effector and target cell stimulation during conjugation by fluorescence polarization.

The aim of the present study was to trace early intracellular changes induced in effector and target cells during their conjugation. This was performed by monitoring the intracellular fluorescein fluorescence polarization (IFFP), using the Cellscan apparatus. This apparatus permits the repetitive spectroscopic measurement of individual selected live cells within a population of many cells, while the location of each cell is known and preserved during the various cell manipulations and/or their suspending medium. Both natural killer (NK) and lymphocyte activated killer (LAK) cells were used as effector cells, while NK-sensitive K562 and NK-resistant Daudi cell lines were used as targets. In this study kinetic IFFP measurements were carried out for a period of approximately 4 h following cell attachment. Within minutes following effector-target conjugation, transient reduction of IFFP was observed consecutively, first in the effector and then in the target cells. A continuous reduction of IFFP occurring only in target cells was also found 50 min following conjugation. No reduction in IFFP was observed using NK- and LAK-resistant target cells. Good correlation was found between early stages of conjugation, as assessed by IFFP, and cytolytic efficiency as assessed by 51chromium release assay. When NK-resistant and LAK-resistant target cells were used, no reduction of IFFP was observed.

Respiratory syncytial virus infection in children with congenital heart disease: a review.

This paper reviews recent changes in morbidity and mortality of respiratory syncytial virus (RSV) infection in infants with congenital heart disease. Mortality since the late 1970s has declined substantially, from approximately 37% to 3%. Although the frequency of admission to intensive care units has declined from approximately 60% to 30%, the frequency for mechanical ventilatory support has not changed significantly. Because mortality dropped prior to the widespread use of ribavirin, it is difficult to ascribe the improvement to this therapy. In infants with congenital heart disease (CHD), nosocomial infection remains a significant problem, accounting for approximately 33% of the RSV cases. Some authors report significant reductions in hospital-acquired RSV by use of gloves and gowns for contacts with infectious cases. Efforts at primary prevention have encountered problems with development of an RSV vaccine. Preliminary data from studies of passive immunization using immune globulins with high RSV antibody titers suggest that this therapy may reduce the severity of RSV infection in infants with serious heart disease.

Ambulatory blood pressure in patients with occult recurrent coarctation of the aorta.

The hypothesis that mild recurrent aortic obstruction produces subtle changes in ambulatory blood pressure was investigated by performing 24-hour monitoring on 11 postoperative coarctation patients. Patients (age 16.1 +/- 2.7 years) were compared with normal controls (age 15.7 +/- 2.5 years, n = 15). Surgery (end-to-end anastomosis) was performed at 6.0 +/- 1.0 years of age. There were no significant differences between patients and controls in terms of baseline blood pressure (right arm 123/78 +/- 4/3 mmHg versus 120/75 +/- 3/2 mmHg) or right leg systolic pressure (125 +/- 6 mmHg versus 123 +/- 4 mmHg). Of the 11 patients 8 had recoarctation by Doppler study (mean gradient 25.3 +/- 2.1 mmHg), 5 of 11 had a postexercise arm-leg pressure difference of > 30 mmHg, and 6 patients had aortic diameters at the site of surgery < 70% of the descending aortic diameter (by magnetic resonance imaging). There were no significant differences between the coarctation and control groups in terms of mean ambulatory systolic (125 +/- 3 mmHg versus 119 +/- 2 mmHg) or diastolic (69 +/- 2 mmHg versus 72 +/- 2 mmHg) pressures throughout the day. However, coarctation patients had a larger number of systolic pressures that exceeded the 95th percentile (18.2 +/- 5.6% versus 6.8 +/- 1.2%). These labile increases in systolic pressure correlated with residual coarctation (r = 0.642, p = 0.003). Ambulatory monitoring is a useful tool for detecting and monitoring subtle abnormalities of blood pressure control after coarctation repair.

Cognitive function in children who receive organ transplantation.

The number of children who undergo cardiac, renal, and liver transplantation continues to increase. This review of the literature describing their cognitive function indicates that deficits ranging from gross IQ delay to subtle neuropsychological dysfunction are present in some of these children both before and after transplantation. However, with the exception of certain definite contributors to vulnerability, such as cyanotic heart disease, early onset of disease, and growth deficits in infancy (particularly in head circumference and height), the correlates of cognitive deficits are not clear. Problems with determining contributing variables include significant heterogeneity within study populations, difficulties obtaining appropriate control groups, and the need for longitudinal, long-term studies. Additional research is required to address limitations of past studies.

Ethnicity and socioeconomic status: impact on the diagnosis of congenital heart disease.

OBJECTIVES: The purpose of this study was to estimate the prevalence rates of specific cardiac defects for three ethnic groups and to determine the effects of ethnicity, family income and household education level on the timing of referral for pediatric cardiac care. BACKGROUND: Previous studies examining ethnic differences in rates of congenital heart disease were based on hospital referrals or were limited to diagnoses made in the 1st year of life. These limitations may lead to potential biases in the ascertainment of cases. The present study is population based and includes patients diagnosed after the 1st year of life. METHODS: Cases of congenital heart disease were enumerated among 379,561 liveborn infants to black, white and Mexican-American residents in Dallas County, Texas. Diagnosis was made on the basis of examination by a pediatric cardiologist, two-dimensional echocardiographic studies, cardiac catheterization or observations at operation or at autopsy. Ethnicity, median family income and household educational level were determined from birth certificate information. RESULTS: White children had higher prevalence rates for aortic stenosis, endocardial cushion defect and ventricular septal defect. Mexican-American children had the lowest rate for hypoplastic left heart syndrome. The median age at referral to a pediatric cardiologist was 1.9 months for blacks, 2.1 months for whites and 2.2 months for Mexican-Americans. Stratifying the cases by median family income and household educational level failed to show any significant relation to age at referral. CONCLUSIONS: Prevalence rates of specific cardiac defects vary among black, white and Mexican-American children, probably reflecting different genetic and environmental backgrounds. The timing of referral for pediatric cardiac care, however, was not related to ethnicity, median family income or household educational level.

Limitations of an oscillometric ambulatory blood pressure monitor in physically active children.

OBJECTIVE: To determine the limitations of an oscillometric device for measuring ambulatory blood pressure in physically active children. DESIGN: Observational descriptive. SUBJECTS: Sixty-one children 4.3 to 18.7 years of age. INTERVENTIONS: Twenty-two high school students wore an ambulatory blood pressure monitor (SpaceLabs model 90202) for 24 hours. A subject-kept activity diary facilitated data interpretation. In 39 additional subjects, measurements were recorded during strenuous exercise, at rest, and after ambulation. Before and after the students wore the unit, oscillometric and auscultatory blood pressure measurements were simultaneously taken to calibrate the ambulatory blood pressure monitor, evaluate its accuracy, and assess deterioration in the calibration of the unit with use. RESULTS: During 24-hour ambulatory monitoring, 29% of the readings were edited because of error codes, primarily because of vibratory interference. At the end of the 24-hour monitoring period, simultaneous pressures by oscillometric and auscultatory techniques agreed within 6 mm Hg in 17 of 18 cases for both systolic and diastolic blood pressures. During exercise, 88.4% of the systolic and diastolic blood pressure readings agreed within 8 mm Hg. In 20 other ambulatory subjects with 80 simultaneous oscillometric and auscultatory measurements, all systolic and 88.5% of the diastolic values agreed within 10%. CONCLUSIONS: These results indicate that the SpaceLabs model 90202 unit provides a reasonably accurate assessment of ambulatory blood pressures in mildly active and inactive children and that blood pressure can be monitored with time.

Ambulatory blood pressure monitoring in hypertensive adolescents.

The purpose of this study was to determine the ability of ambulatory blood pressure monitoring to identify youths with chronic blood pressure elevation. Nineteen adolescent boys were studied, ten had 5-year average systolic or diastolic pressures above the 95th percentile, nine had normal pressure. A Del Mar Avionics Pressurometer III system recorded an average of 121 readings on each subject. The coefficients of variation for pressure were similar for hypertensive and normotensive individuals. During classes, eight of the ten hypertensive youths had elevated pressures in over half of the measurements. Also during these classes eight of ten hypertensive boys had average systolic or diastolic pressure above the 95th percentile, whereas only one of nine normotensive boys had average pressures above this level. We suggest that schooltime ambulatory pressures may be most useful in classifying the blood pressure trend in a youth.

Trends in congenital heart disease in Dallas County births. 1971-1984.

To examine the changes in birth cohort prevalence rates and severity of congenital heart disease, we studied children with congenital heart disease born to blacks, whites, and Mexican-Americans in Dallas County from 1971 through 1984. Diagnoses were made by pediatric cardiologists' clinical evaluations, echocardiography, catheterization, surgery, or autopsy. During this study period, 2,509 of 379,561 liveborn infants were diagnosed, a prevalence rate of 6.6/1000. The rates for whites was significantly higher than for blacks or Mexican-Americans--7.2/1,000, 5.6/1,000, and 5.9/1,000, respectively. The rate for severe cases requiring cardiac catheterization or surgery or undergoing autopsy was 3.1/1,000 and did not differ among the three groups. The time trend for rates of congenital heart disease suggested an apparent increase in prevalence rate during the 1970s; however, the prevalence rate of severe forms remained relatively stable. This indicates that the apparent rise in prevalence could be accounted for by an increase in detection of mild cases. These findings were interpreted as reflecting a greater tendency for pediatricians to refer asymptomatic children with significant heart murmurs to a pediatric cardiologist.