Whole-body lymphangioscintigraphy and SPECT/CT in children with lymphatic complications after surgery for complex congenital heart disease.

The number of patients surviving repair of complex congenital heart disease (CCHD) has increased due to improved surgical techniques, post operative management and outpatient care. Likewise, this growing patient population has demonstrated an increasing number and complexity of complications involving the lymphatic system. To evaluate the peripheral and central lymphatic system, whole-body lymphangioscintigraphy (LAS) is considered as the initial imaging evaluation of choice. To date, very few publications exist on the value of lymphatic imaging techniques in infants and small children with lymphatic complications following surgery for congenital heart disease. A retrospective review of medical records from 2008 to 2018 was performed for pediatric patients referred for lymphatic complications after CCHD surgery at an academic medical center. LAS and SPECT/CT was performed using intradermal bipedal injections of Tc 99m labeled filtered sulfur colloid, and in some patients also bilateral hand injections, followed by dynamic imaging and whole- body planar imaging typically up to 180 minutes post injection. Clinical decision making and outcomes were recorded. LAS and SPECT/CT were performed without complication in pediatric patients with prior surgery for CCHD. LAS successfully localized various lymphatic abnormalities such as lymphatic obstruction, reflux, and leaks, which were further delineated by SPECT/CT. LAS findings directed further evaluation with more definitive studies, management and prognosis. Five of the ten patients had follow up outcome data - 2 years and up to 10 years. LAS and SPECT/CT are safe and effective techniques for the initial evaluation of lymphatic abnormalities in pediatric patients with CCHD. LAS, particularly with further 3D localization by SPECT/CT, provides functional imaging of peripheral and central lymphatic flow and thus provides guidance for medical therapy, non operative interventional management, and surgical therapy for these diverse, debilitating, and often life threatening disorders.

Successful treatment of plastic bronchitis with low fat diet and subsequent thoracic duct ligation in child with fontan physiology.

Plastic bronchitis is a rare condition characterized by the formation and expectoration of long, branching bronchial casts that develop in the tracheobronchial tree and cause airway obstruction. Plastic bronchitis has become increasingly recognized as a feared complication of the Fontan operation with a mortality of up to 50%. We report an 11 year old boy who developed severe plastic bronchitis following Fontan repair and the successful long-term control of cast formation utilizing a low-fat diet and subsequent thoracic duct ligation.

The dicrotic pulse: a common, non-ominous finding after the Ross operation.

We noted a dicrotic pulse in several patients following a Ross operation. Although the etiology of this unique arterial waveform is not completely understood, it has been reported as a sign of low cardiac output and a poor prognosis. We reviewed preoperative echocardiograms and postoperative radial arterial pressure tracings in 33 patients who underwent a Ross procedure between 2000 and 2004. We found a dicrotic pulse to occur commonly (20/33; 61%) following a Ross operation. Moderate to severe preoperative aortic insufficiency was present in 19/20 patients (95%) in whom a dicrotic pulse was noted and in only 3/13 (23%) who did not exhibit a postoperative dicrotic pulse (p<0.001). A dicrotic pulse was not associated with an increased use of vasoactive infusions or longer hospitalization following the Ross operation. The dicrotic pulse should be recognized as a common postoperative finding in Ross patients that does not herald a delayed postoperative convalescence. The mechanism for a dicrotic pulse in these patients is speculative but may result from changes in vascular compliance secondary to chronic aortic insufficiency.

Resolution of plastic bronchitis with atrial pacing in a patient with fontan physiology.

We describe a 5-year-old boy with Fontan physiology and a ventricular pacemaker who developed severe plastic bronchitis. Evaluation by cardiac catheterization revealed profoundly altered hemodynamics, which improved with atrial pacing. Following implantation of an atrial pacemaker, which restored atrioventricular (A-V) synchrony, the patient's hemodynamics greatly improved and his plastic bronchitis resolved.

Development of a PCR assay for detection of the oyster pathogen Bonamia ostreae and support for its inclusion in the Haplosporidia.

The development of diagnostic assays more sensitive and specific than traditional histological techniques is important for the management of bonamiasis in flat oysters Ostrea edulis. A specific polymerase chain reaction (PCR) protocol was developed for the detection of very small amounts of Bonamia ostreae (Pichot et al. 1980) ribosomal DNA (rDNA) in bulk DNA from oyster gill and hemolymph. The presence of a 760 bp PCR amplification product corresponded with B. ostreae infections determined cytologically in 185 oysters from Ireland, Spain, and the USA. All (100%) 'heavily' and 'moderately' infected oysters, 86.7 % of the 'lightly' infected oysters, and 66.7 % of the 'scarcely' infected oysters were confirmed to be infected using the PCR. In addition, 37.9% of the oysters in which B. ostreae was not detected using cytology were positive using the PCR. Sampling error and the subjectivity of cytological diagnoses are the likely sources of disagreement between diagnostic methods in oysters with very light infections. The PCR assay developed here is more sensitive and less ambiguous than standard histological and cytological techniques. Phylogenetic analysis of DNA sequence data confirmed B. ostreae to be a member of the Haplosporidia.

Additional evidence that juvenile oyster disease is caused by a member of the Roseobacter group and colonization of nonaffected animals by Stappia stellulata-like strains.

Juvenile oyster disease (JOD) causes significant annual mortalities of hatchery-produced Eastern oysters, Crassostrea virginica, cultured in the Northeast. We have reported that a novel species of the alpha-proteobacteria Roseobacter group (designated CVSP) was numerically dominant in JOD-affected animals sampled during the 1997 epizootic on the Damariscotta River, Maine. In this study we report the isolation of CVSP bacteria from JOD-affected oysters during three separate epizootics in 1998. These bacteria were not detected in nonaffected oysters at the enzootic site, nor in animals raised at a JOD-free site. Animals raised at the JOD enzootic site that were unaffected by JOD were stably and persistently colonized by Stappia stellulata-like strains. These isolates (designated M1) inhibited the growth of CVSP bacteria in a disk-diffusion assay and thus may have prevented colonization of these animals by CVSP bacteria in situ. Laboratory-maintained C. virginica injected with CVSP bacteria experienced statistically significant elevated mortalities compared to controls, and CVSP bacteria were recovered from these animals during the mortality events. Together, these results provide additional evidence that CVSP bacteria are the etiological agent of JOD. Further, there are no other descriptions of specific marine alpha-proteobacteria that have been successfully cultivated from a defined animal host. Thus, this system presents an opportunity to investigate both bacterial and host factors involved in the establishment of such associations and the role of the invertebrate host in the ecology of these marine alpha-proteobacteria.

Use of antibacterial agents To elucidate the etiology of juvenile oyster disease (JOD) in Crassostrea virginica and numerical dominance of an alpha-proteobacterium in JOD-affected animals.

Since 1988, juvenile oyster disease (JOD) has resulted in high seasonal losses of cultured Eastern oysters (Crassostrea virginica) in the Northeast. Although the cause of JOD remains unknown, most evidence is consistent with either a bacterial or a protistan etiology. For the purpose of discerning between these hypotheses, the antibacterial antibiotics norfloxacin and sulfadimethoxine-ormetoprim (Romet-B) were tested for the ability to delay the onset of JOD mortality and/or reduce the JOD mortality of cultured juvenile C. virginica. Hatchery-produced C. virginica seed were exposed in triplicate groups of 3,000 animals each to either norfloxacin, sulfadimethoxine-ormetoprim, or filter-sterilized seawater (FSSW) and deployed in floating trays on the Damariscotta River of Maine on 17 July 1997. Each week thereafter, a subset of animals from each group was reexposed to the assigned treatment. Repeated immersion in either a sulfadimethoxine-ormetoprim or a norfloxacin solution resulted in a delay in the onset of JOD mortality in treated animals and reduced weekly mortality rates. Weekly treatments with either norfloxacin or sulfadimethoxine-ormetoprim also resulted in a statistically significant reduction in cumulative mortality (55 and 67% respectively) compared to animals treated weekly with FSSW (81%) or those that had received only a single treatment with either norfloxacin, sulfadimethoxine-ormetoprim, or FSSW (77, 84, and 82%, respectively). Bacteriological analyses revealed a numerically dominant bacterium in those animals with obvious signs of JOD. Sequence analysis of the 16S rRNA gene from these bacteria indicates that they are a previously undescribed species of marine alpha-proteobacteria.

Haplosporidium nelsoni (MSX) epizootic in the Piscataqua River Estuary (Maine/New Hampshire, USA.).

A major epizootic of Haplosporidium nelsoni occurred during 1995 in the Piscataqua River Estuary, near the northern distributional limit of the parasite. Prevalence of H. nelsoni in samples of oysters, Crassostrea virginica, collected between 5 September and 18 December 1995 from sites in Maine and New Hampshire ranged from 15 to 81%, with up to 50% systemic infections. Associated proportions of dead oysters ranged from 25 to 83%. The epizootic was most likely triggered by unusually warm and dry climatic conditions in 1995 that resulted in a temperature and salinity regime in the estuary that favored the parasite.

Effects of gonadal neoplasms on oogenesis in softshell clams, Mya arenaria.

The average prevalence of gonadal neoplasms in softshell clams, Mya arenaria, from Whiting Bay, Washington County, Maine, was 19.4% in 1994. Monthly prevalences ranged from 10 to 26.7%. Neoplasms ranged in intensity from few, small foci of undifferentiated germ cells (Stage 1), to 50-100% of gonadal follicles being involved (Stage 2), to invasion and metastasis with loss of tissue architecture (Stage 3), indicating that the disease is progressive and lethal. There was no relationship (P > 0.05) between prevalence of neoplasms and clam size between 45.7 and 60.7 mm mean shell length. Clams of both sexes were affected, but females were significantly more likely (P

Texture analysis of protein distribution images to find differences due to aging and superfusion.

Many pathologies are age-related, e.g., cardiovascular disease generally occurs in midlife and cancer later in life. This suggests that aging predisposes the body to pathology. Plasma protein spatial distribution images of rat mesentery extracellular matrix (ECM) show texture due to the ECM structures, and there is an age-related decrease in tissue protein that may be related to matrix structure changes. The objective of this study was to compare changes in protein image texture under two conditions: superfusion with normal saline solution and aging. The decrease in soluble protein concentration during superfusion is a washout process, while the mechanism for the age-related decrease in tissue protein is unknown. Therefore, effects of aging and superfusion on tissue protein image texture were compared. Spatial co-occurrence matrix and Fourier analysis techniques have been used for texture evaluation. Superfused images showed a more uniform protein texture. There were gradual age-related changes in image texture parameters. Entropy increased with age from 140 to 630 days, indicating that protein distribution became more disorganized. The results suggest that changes in protein image texture are due to age-related alterations in matrix structure because removing only protein by superfusion had opposite effects on texture parameters.

Age-related changes in albumin-excluded volume fraction.

The space available to large macromolecules, such as albumin and globulin, is less than the total interstitial fluid volume due to the dense matrix formed by the interstitial ground substance. Changes in excluded volume are likely to indicate changes in the composition of the matrix. Sprague-Dawley rats were anesthetized with sodium pentobarbital. Serum, mesenteric tissue, and peritoneal fluid samples were obtained. Albumin contents were determined by microrod electrophoresis. Serum and mesenteric tissue chloride concentrations were measured by the coulometric-amperometric method. Serum and mesenteric tissue sample chloride concentrations were not significantly different, suggesting that this loose connective tissue is composed almost entirely of extracellular matrix. Matrix hydration decreased with a regression slope of -0.014 (microgram tissue water/microgram tissue dry wt)/10 days. Serum and tissue albumin concentrations decreased between 210 and 630 days of age. Mesenteric loose connective tissue albumin-excluded volume fraction increased by 80% over this age range. The increase could not be accounted for by dehydration alone, suggesting that the increase in excluded volume fraction for albumin is also due to changes in tissue glycosaminoglycans or collagen.

Age-related changes in perimicrovascular protein distribution.

The diffusion hypothesis for physiological aging proposes that an increase in interstitial matrix fiber-to-gel ratio causes a decrease in nutrient diffusion to the cells. This hypothesis predicts a decrease in interstitial matrix protein with age. The objective was to test this hypothesis by determining age-related changes in plasma protein distribution in perimicrovascular and distal regions of rat mesentery interstitial matrix. Rats that were 77, 140, 210, 315, 455, and 630 days old were anesthetized with pentobarbital sodium, and a mesenteric loop was exteriorized. Intravital video microspectrophotometry was performed using wavelengths of 280, 320, and 700 nm. Perimicrovascular protein concentrations from the protein absorbance images were used to obtain the histogram, mean, and skewness of the proximal and distal protein concentration distributions. An exponential gradient model was also used to obtain the proximal and distal protein concentrations and gradient decay constants. Proximal protein concentration increased from 77- to 140-day-old rat and then decreased gradually through 210-, 315-, 455-, and 630-day-old rats. Distal concentration decreased gradually from 140- to 630-day-old rats. There was an increase in positive skewness of the proximal protein distributions from 140- through 630-day-old rats. We found an age-related decrease in perimicrovascular protein and propose that this is due to a decrease in protein permeability with age. The results support the diffusion theory of aging.

Age-related changes in rat interstitial matrix hydration and serum proteins.

Measurements of effects of age-related changes on loose connective tissue protein concentrations and water are lacking. Tissue hydration is an important determinant of tissue protein diffusion coefficients and hydraulic conductivity. Sprague-Dawley rats (ages 11, 30, 57, and 89 wk) were anesthetized. Blood and mesenteric tissue samples were taken. Tissue water content was determined by microgravimetric technique. Protein content was determined by electrophoresis. Tissue hydration decreased 18% between 11 and 89 wk with one-half the fall occurring between 57 and 89 wk. Tissue albumin concentration decreased 37% between 11 and 89 wk. Serum albumin increased 22% between 11 and 30 wk and then decreased by 37% to 1.4 g/dl in 89-wk rats. Serum total protein concentration increased by 13% between 11 and 89 wk, whereas tissue total protein concentration decreased 30% with age to 1.9 g/dl at 89 wk. The results suggest that age-related tissue dehydration is due, in part, to a shift in the colloid osmotic pressure components of Starling equilibrium favoring fluid reabsorption.

Changes in perimicrovascular protein spatial distribution due to superfusate.

OBJECTIVE: To determine superfusate-induced changes in the distribution of plasma proteins in the perimicrovascular interstitial matrix. METHODS: Rats were anesthetized with sodium pentobarbital and a mesenteric loop was exteriorized. Intravital video microspectrophotometry was performed using wavelengths of 280, 320, and 700 nm. The images were analyzed to give protein and collagen spatial distributions in vascular regions of rat mesenteric tissue. Perimicrovascular protein concentrations were fitted to an exponential decay model ci + cr exp (-x/k), where ci is distal protein concentration, ci + cr is the protein concentration proximal to the vessel, x is the distance from the vessel wall, and k is the decay constant indicating protein gradient slope. RESULTS: Before superfusion with 0.5-ml normal saline, ci = 1.45 +/- 0.13 g/dl, ci + cr = 4.56 +/- 0.23 g/dl. After the first superfusion, ci decreased (p < 0.01) to 0.53 +/- 0.06 g/dl; following a second superfusion, cr decreased to 0.4 +/- 0.03 g/dl; an additional final superfusion caused a further decrease to 0.33 +/- 0.02 g/dl. ci + cr also decreased significantly during repeated superfusions to 2.92 +/- 0.15, 2.35 +/- 0.25, and 2.1 +/- 0.12 g/dl, respectively. CONCLUSIONS: Superfusion produced changes in perivascular and distal interstitial matrix protein distribution. Protein concentration proximal to the microvessel remained higher than distal concentrations. This could be due to increased gel concentrations inhibiting protein mobility.

Changes in rat mesentery interstitial matrix due to superfusate.

Animal preparations for microscopy often require a superfusate solution to cover surgically exposed tissue. There are few, if any, data concerning the effects of this solution on extravascular protein concentration and hydration. The effect of superfusion on mesenteric tissue in anesthetized male Sprague-Dawley rats was studied. Tissue samples were taken from nonsuperfused and superfused tissue and analyzed for hydration, albumin, and transferrin content. The mesenteric tissue interstitial matrix was rapidly altered by normal saline superfusate. After superfusion, there was a decrease (P < 0.01) in tissue albumin concentration from 1.17 +/- 0.27 to 0.10 +/- 0.08 g/dl (n = 9). Tissue hydration increased from 4.98 +/- 0.8 micrograms water/microgram dry wt in controls to 7.38 +/- 1.2 micrograms water/micrograms dry wt after superfusion. When a range of superfusate albumin concentrations was used (0, 1, 2, and 3 g/dl), tissue albumin concentration changed 0.59 +/- 0.09 g/dl for each gram per deciliter change in superfusate concentration (P < 0.0001). The large changes in interstitial matrix protein content and hydration suggest that superfusate solution effects need to be considered in microvascular protein transport experiments.

Bromcresol green assay is nonspecific for rat plasma albumin.

Bromcresol green (BCG) assay has been used in microvascular studies to determine albumin in rat plasma. The purpose of this study was to demonstrate that BCG overestimates rat plasma albumin partly because BCG binds to transferrin, a beta-globulin. The light absorbance of a transferrin-BCG reagent solution is shown to increase with time; appreciable binding occurs within a few seconds. Pure transferrin produced BCG assay results (P less than 0.001) that could be expressed as pseudoalbumin concentrations. Albumin and transferrin solutions of equal concentrations were mixed in equal parts; a plot of albumin concentration determined by BCG vs. actual albumin concentration had a slope greater than the expected value of 1.0 (P less than 0.001). Plasma samples were obtained from six rats and assayed for albumin. Electrophoresis yielded a plasma albumin of 2.97 +/- 0.11 g/dl, whereas BCG assay yielded a significantly (P less than 0.01) greater value of 3.58 +/- 0.07 g/dl. We conclude that BCG assay estimates of albumin-to-globulin ratio (A/G), in which G is determined from the difference between total protein and albumin, are especially subject to error.

Genetic Variation in the Timing of Gonadal Maturation and Spawning of the Eastern Oyster, Crassostrea virginica (Gmelin).

The gonadal cycles of four groups of eastern oysters, Crassostrea virginica (Gmelin), including native stocks collected that year and inbred strains (reared in Delaware Bay for 5-6 generations) from both Long Island Sound and Delaware Bay, were compared in Delaware Bay in 1987. Inbred strains resembled their respective native stocks; both Long Island groups initiated gonadal development and spawning about one month earlier and spawned over a shorter duration than both Delaware Bay groups. Analysis of covariance revealed that the effect of time on gonadal development was statistically different (P ≤ 0.05) for all between-location group comparisons, but not for the two within-location comparisons. Thus, after six generations of inbreeding in Delaware Bay, Long Island oysters maintained their characteristic pattern of gonadal development and spawning, indicating the existence of genetically different environmental requirements for gonadal maturation between the two locations.

Thermal effects of MR imaging: worst-case studies on sheep.

The objective of this study was to provide a worst-case estimate of thermal effects of MR imaging by subjecting anesthetized unshorn sheep to power deposition at specific absorption rates (SARs) well above approved standards for periods of time in excess of normal clinical imaging protocols. A control period with no RF power was followed by 20-105 min of RF power application. Afterward, there was a 20-min or longer recovery period with no RF power. Eight sheep were given whole-body RF exposure (1.5- to 4-W/kg SAR) while rectal and skin temperatures were monitored. Four sheep were subjected to 4-W/kg head scans for an average of 75 min while temperatures of the cornea, vitreous humor, head skin, jugular vein, and rectum were measured. In head scanning experiments, skin and eye temperatures increased about 1.5 degrees C. Jugular vein temperature rose a maximum of 0.4 degrees C after an average exposure of 75 min. In whole-body exposures, elevation of rectal temperature was correlated with energy input. Deep-body temperature rises in excess of 2.0 degrees C were attained for 4-W/kg whole-body exposure periods greater than 82 min. Animals exposed for 40 min to 4 W/kg in either body coil (three sheep) or head coil (two sheep) were recovered and observed to be in good health for 10 weeks; no cataracts were found. MR power deposition at SAR levels well above typical clinical imaging protocols caused body temperature to increase. For exposure periods in excess of standard clinical imaging protocols the temperature increase was insufficient to cause adverse thermal effects. Studies in healthy humans are needed to determine whether enhanced heat-loss effector mechanisms are likely to cause deep-body temperatures to plateau at an acceptable level, and to elucidate mechanisms that determine subcutaneous temperature.

Comparative analysis of protein content in rat mesenteric tissue, peritoneal fluid, and plasma.

Albumin, transferrin, and total protein concentrations were measured in the mesenteric tissue, peritoneal fluid, and plasma of 12 ketamine-Nembutal-anesthetized Sprague-Dawley rats. Tissue samples were obtained with an 8-mm trephine; tissue water content was determined by a microgravimetric method to be 5.2 +/- 0.3 microgram water/microgram dry wt. Peritoneal fluid was collected by capillary action in hematocrit tubes, and blood samples were taken from a femoral artery catheter. Total protein concentrations of plasma (5.8 +/- 0.3 g/dl) and peritoneal fluid (2.6 +/- 0.1 g/dl) were determined by Lowry assay. Ratios of peritoneal fluid and tissue densitogram areas to plasma area were used to calculate total protein content of peritoneal fluid (2.5 +/- 0.1 g/dl) and tissue (1.8 +/- 0.2 g/dl). Albumin concentrations were 1.1 +/- 0.1 g/dl for tissue, 1.4 +/- 0.1 g/dl for peritoneal fluid, and 2.8 +/- 0.1 g/dl for plasma. Transferrin concentrations were 0.09 +/- 0.01 g/dl for tissue, 0.13 +/- 0.01 g/dl for peritoneal fluid, and 0.28 +/- 0.01 g/dl for plasma. Peritoneal fluid protein concentrations were similar to values found for lymph in previous studies. Protein concentration in the tissue buttons was significantly less than that of peritoneal fluid. This contradicts the widely held assumption that the protein concentration of fluid outside the matrix is representative of a well-mixed interstitial matrix fluid protein concentration.

Spatial distribution of protein in interstitial matrix of rat mesenteric tissue.

Interstitial matrix protein has been measured by collecting fluid from implanted capsules and wicks. If the interstitial matrix is well mixed, then these measurements provide an accurate indication of perivascular protein concentration. However, there is increasing evidence suggesting that interstitial protein distribution is nonuniform. To test this hypothesis, a combined light-scattering and protein absorbance image of mesenteric tissue in a pentobarbital-anesthetized rat was obtained by using an ultraviolet-sensitive television camera to perform videomicrospectrophotometry. In avascular tissue regions protein was organized into periodic nonuniform ridges and valleys. On a smaller scale, the protein along a ridge was organized into clusters. Similar clusters were seen along the wall of small tortuous microvessels and were accompanied by radial gradients in protein distribution. High protein concentration tunnels were occasionally seen extending from a microvessel into the interstitial matrix. A two-way analysis of variance shows that macromolecular spatial distribution is nonuniform (P less than 0.001). Thus the hypothesis that the extracellular matrix is well mixed fails for macromolecules in rat mesenteric loose connective tissue.