Biomedical subjects
Publications and source records attributed to M L Epstein.
There is little experience using beta-blocker therapy for the management of congestive heart failure (CHF) in children. We present the case of a child with idiopathic dilated cardiomyopathy and CHF, referred for cardiac transplantation, in whom carvedilol reversed elevated pulmonary vascular resistance.
The enteric nervous system (ENS) develops from neural crest cells that enter the gut, migrate, proliferate, and differentiate into neurons and glia. The growth factor glial-derived neurotrophic factor (GDNF) stimulates the proliferation and survival of enteric crest-derived cells. We investigated the intracellular signaling pathways activated by GDNF and their involvement in proliferation. We found that GDNF stimulates the phosphorylation of both the PI 3-kinase downstream substrate Akt and the MAP kinase substrate ERK in cultures of immunoaffinity-purified embryonic avian enteric crest-derived cells. The selective PI 3-kinase inhibitor LY-294002 blocked GDNF-stimulated Akt phosphorylation in purified crest cells, and reduced proliferation in cultures of dissociated quail gut. The ERK kinase (MEK) inhibitors PD 98059 and UO126 did not reduce GDNF-stimulated proliferation, although PD 98059 blocked GDNF-stimulated phosphorylation of ERK. We conclude that the PI 3-kinase pathway is necessary for the GDNF-stimulated proliferation of enteric neuroblasts.
Performance on two multiple-choice testing procedures was examined during unit tests and a final examination. The Immediate Feedback Assessment Technique provided immediate response feedback in an answer-until-correct style of responding. The testing format which served as a point of comparison was the Scantron form. One format was completed by students in introductory psychology courses during unit tests whereas all students used the Scantron form on the final examination. Students tested with Immediate Feedback forms on the unit tests correctly answered more of the final examination questions which were repeated from earlier unit tests than did students tested with Scantron forms. Also, students tested with Immediate Feedback forms correctly answered more final examination questions previously answered incorrectly on the unit tests than did students tested previously with Scantron forms.
OBJECTIVES: This study was undertaken to determine the potential role of P wave signal-averaged electrocardiogram (PSAECG) for risk assessment of atrial tachyarrhythmias (ATs) in patients after Fontan operation. BACKGROUND: Onset of atrial flutter/fibrillation (AFF) in patients who have undergone Fontan operation for univentricular hearts constitutes an unfavorable clinical event associated with a high risk of cardiovascular complications. There is no data available on PSAECG in postoperative Fontan patients to predict potential susceptibility to ATs. METHODS: Twenty-four post-Fontan patients and 15 age-matched healthy subjects were prospectively studied with PSAECG, and the following measurements were made: filtered P wave duration (FPWD), P wave vector integrals (PINTs), root-mean-square voltage for the initial 30 ms (RMSi30), and duration of persistent amplitude signals <4 microV from the beginning of the P wave (Di4). RESULTS: The FPWDs were significantly longer in the study group patients with ATs when compared with the study group patients without ATs (p<0.01) and compared with the controls (p < 0.001). An FPWD cut point of 135 ms resulted in a sensitivity of 71% and a specificity of 81% in differentiating patients with ATs from patients without ATs among the postoperative Fontan patients. The PINT was significantly greater in Fontan patients with AFF and also without AFF when compared with controls (p<0.01, p<0.05, respectively). The RMSi30 and the Di4 were not significantly different between study and control groups. CONCLUSIONS: Signal-averaged P wave duration is significantly prolonged in postoperative Fontan patients. A prolonged signal-averaged P wave duration may be an effective noninvasive marker to predict risk of development of ATs in this patient group.
The formation of the enteric nervous system (ENS) from neural crest-derived cell precursors requires the growth factor glial cell line-derived neurotrophic factor (GDNF) and the receptors Ret and GDNF family receptor alpha 1 (GFRalpha1). We investigated the location(s), the timing, and the extent to which these GDNF receptors appear in the population of crest-derived precursors that form the avian ENS using immunohistochemistry and in situ hybridization. Sections and whole mounts of embryonic chick gastrointestinal tract were costained with antibodies to the receptors and to HNK-1, a marker for crest-derived cells. Neural crest-derived precursors migrate through the primitive esophagus to colonize the gizzard where an extensive cellular network forms. Ret-immunoreactivity (ir) was found in a network of cells in the gizzard at embryonic day (E)3.5. As development proceeded, Ret-immunoreactive cells appeared at progressively more caudal positions and were present in the colon at E7.5. Costaining with Ret and HNK-1 was performed to determine the number of Ret-immunoreactive cells in the crest-derived population. Ret appeared in some HNK-1 cells in the esophagus and gizzard at embryonic day (E)3.5. During development, the number of crest cells with Ret increased in the ganglia of the gizzard and small intestine. GFRalpha1-ir was also found in HNK-1 cells in the esophagus at E3.5 but did not appear in the gizzard until E4.5. Surprisingly, the colonizing vanguard of crest-derived cells lacked both Ret- and GFRalpha-ir. Between E4.5 and E6.5, the fraction of HNK-1-positive cells expressing GFRalpha1 increased considerably in the foregut. Ret and GFRalpha1 were coexpressed in many cells at E6.5, and the number of such cells increased as development progressed. In the adult, GFRalpha1 and Ret were found in the neuropil of enteric ganglia. We conclude that the population of cells expressing the receptors increases during development and persists in the adult, findings that support a neurotrophic role for GDNF in the formation and maintenance of the avian ENS.
This study evaluated changes in neoaortic root geometry in patients who underwent the Ross procedure. Serial postoperative echocardiographic measurements of the neoaortic root indexed to the square root of body surface area (centimeters divided by meters) were obtained from 30 patients (age range 3.1 to 31.4 years) and compared with paired preoperative and immediate postoperative values. Normal aortic root diameter Z scores were derived from root dimensions obtained from 217 healthy controls. Compared with preoperative values, an immediate stretch of the neoaortic versus pulmonary root (annulus and sinuses of valsalva) was observed at a mean follow-up period of 1 week. Additional aortic annular dilation from baseline prehospital discharge values was observed at 2 to 12 months (baseline vs follow-up annulus Z score: 1.4 vs 2.6, p <0.01, n = 16) and at 16 to 33 months follow-up (0.8 vs 2.0, p <0.05, n = 12). In a similar fashion, there was additional enlargement of the aortic sinus from its stretched state at hospital discharge at 2 to 12 months (baseline vs follow-up sinus Z score: 2.0 vs 3.3, p <0.01, n = 17) and at 16 to 33 months (1.7 vs 3.0, p <0.01, n = 13). There were no differences in root size between 2 to 12 and 16 to 33 months after surgery. There was a decrease in left ventricular size with no alteration in blood pressure or degree of aortic valve regurgitation. Thus, aortic root dilation occurs up to the first year after the Ross procedure but does not appear to progress beyond this time.
Vitamin A is required for reproduction and normal embryonic development. We have determined that all-trans-retinoic acid (atRA) can support development of the mammalian embryo to parturition in vitamin A-deficient (VAD) rats. At embryonic day (E) 0.5, VAD dams were fed purified diets containing either 12 micrograms of atRA per g of diet (230 micrograms per rat per day) or 250 micrograms of atRA per g of diet (4.5 mg per rat per day) or were fed the purified diet supplemented with a source of retinol (100 units of retinyl palmitate per day). An additional group was fed both 250 micrograms of atRA per g of diet in combination with retinyl palmitate. Embryonic survival to E12.5 was similar for all groups. However, embryonic development in the group fed 12 micrograms of atRA per g of diet was grossly abnormal. The most notable defects were in the region of the hindbrain, which included a loss of posterior cranial nerves (IX, X, XI, and XII) and postotic pharyngeal arches as well as the presence of ectopic otic vesicles and a swollen anterior cardinal vein. All embryonic abnormalities at E12.5 were prevented by feeding pharmacological amounts of atRA (250 micrograms/g diet) or by supplementation with retinyl palmitate. Embryos from VAD dams receiving 12 micrograms of atRA per g of diet were resorbed by E18.5, whereas those in the group fed 250 micrograms of atRA per g of diet survived to parturition but died shortly thereafter. Equivalent results were obtained by using commercial grade atRA or atRA that had been purified to eliminate any potential contamination by neutral retinoids, such as retinol. Thus, 250 micrograms of atRA per g of diet fed to VAD dams (approximately 4.5 mg per rat per day) can prevent the death of embryos at midgestation and prevents the early embryonic abnormalities that arise when VAD dams are fed insufficient amounts of atRA.
Mutations in HERG are associated with human chromosome 7-linked congenital long QT (LQT-2) syndrome. We used electrophysiological, biochemical, and immunohistochemical methods to study the molecular mechanisms of HERG channel dysfunction caused by LQT-2 mutations. Wild type HERG and LQT-2 mutations were studied by stable and transient expression in HEK 293 cells. We found that some mutations (Y611H and V822M) caused defects in biosynthetic processing of HERG channels with the protein retained in the endoplasmic reticulum. Other mutations (I593R and G628S) were processed similarly to wild type HERG protein, but these mutations did not produce functional channels. In contrast, the T474I mutation expressed HERG current but with altered gating properties. These findings suggest that the loss of HERG channel function in LQT-2 mutations is caused by multiple mechanisms including abnormal channel processing, the generation of nonfunctional channels, and altered channel gating.
Glial derived neurotrophic factor (GDNF) is essential for the development of the enteric nervous system (ENS). Although previous work has measured GDNF mRNA levels, little is known about the concentration of GDNF protein produced in developing or adult tissues. The aim of this study was to quantitate the concentration of GDNF protein in various tissues of the developing and adult rat and in adult human gut. A two site antibody immunoassay was used to quantitate GDNF using recombinant rat GDNF as a standard. In the adult rat gastrointestinal tract the intestine contained the highest concentration of GDNF while the stomach and esophagus have the lowest concentrations. The isolated muscular wall of the intestine has approximately four times the GDNF concentration of the intact intestine. Other tissues with smooth muscle such as the aorta and urinary bladder contain moderate GDNF concentrations. In contrast, GDNF is barely detectable in the adult kidney and liver. High concentrations of GDNF were also detected in human colon and jejunum. As development proceeds in the rat, there is a tendency for the concentration of GDNF to increase in the intestine but decrease in other tissues. Treatment of the jejunum with the cationic surfactant benzyldimethyltetradecylammonium chloride (BAC) results in an increase in the number of smooth muscle cells, a decrease in myenteric neurons, and an increase in the concentration of GDNF in homogenates of intestine. The observations that GDNF concentrations are high in the adult intestine suggest that this growth factor may be important for the maintenance of the adult ENS.
M-mode and Doppler echocardiographic analyses of left ventricular (LV) shortening and filling were performed in 50 patients who underwent coarctectomy (median follow-up 9.5 years) and in 16 athletes in a control group before an exercise stress test with upright bicycle ergometry was performed. Thirty-two of 50 patients and 18 of 50 patients had a normotensive and hypertensive response to exercise, respectively. Preexercise echocardiographic data were compared among the control, normotensive, and hypertensive patient groups. LV peak filling rates (dD/dt, diastole) were increased in the hypertensive group (18.3 +/- 3.5) compared with those in the normotensive group (14.4 +/- 3.2; p < 0.001) and the control group (13.6 +/- 2.8; p < 0.001). LV shortening was enhanced in the coarctectomy group compared with that in the control group. A higher aortic isthmus Doppler gradient at peak exercise was not found in the hypertensive group compared with that in the normotensive group. Therefore patients with successful coarctectomy in childhood have enhanced LV shortening and relaxation at rest. Demonstration of enhanced LV peak filling rates may help identify patients at risk for exercise-induced hypertension.
PURPOSE: To determine the incidence of clinical cardiotoxicity from anthracycline chemotherapy in children with cancer and to identify associated risk factors. PATIENTS AND METHODS: The study population consisted of 6,493 children with cancer who had received anthracycline chemotherapy on Pediatric Oncology Group (POG) protocols from 1974 to 1990. Cardiotoxicity, defined as congestive heart failure not due to other causes, abnormal measurements of cardiac function that prompted discontinuation of therapy, or sudden death from presumed cardiac causes, was determined by a review of protocol records. RESULTS: Cardiotoxicity was confirmed in 106 patients (1.6%): 58 had congestive heart failure, 43 had changes in measures of cardiac function that prompted the discontinuation of therapy, and five died suddenly from presumed cardiac causes. In a multivariate analysis, factors that contributed to the relative risk (RR) of toxicity were a cumulative anthracycline dose > or = 550 mg/m2 of body-surface area (RR = 5.2), maximal dose > or = 50 mg/m2 (RR = 2.8), female sex (RR = 1.9), black race (RR = 1.7), presence of trisomy 21 (RR = 3.4), and exposure to amsacrine (RR = 2.6). Cardiotoxicity within 1 year after the completion of anthracycline treatment (early cardiotoxicity) represented 89.5% of all cases. CONCLUSION: Early clinical cardiotoxicity in children treated with anthracycline is rare. A high maximal dose, or cumulative dose of anthracycline, female sex, black race, presence of trisomy 21, and treatment with amsacrine increase the risk for anthracycline-associated cardiotoxicity.
Mammalian pre-pro-vasoactive intestinal peptide (pre-proVIP) gives rise to the neuropeptides vasoactive intestinal peptide (VIP) and peptide histidine isoleucine amide (PHI). The cDNA encoding chicken VIP was cloned and sequenced. The region of chicken pre-proVIP homologous to the mammalian PHI region is not followed by an amidation signal. This unusual feature suggests that processing of the precursor may be different in the chicken.
The enteric nervous system is formed from neural crest-derived cells. These cells enter the gut, migrate, proliferate, and ultimately differentiate into neurons and glia. We have used a human anti-neuronal autoantibody (ANNA-1), which recognizes neuron-specific RNA-binding proteins of the Hu family as an early marker of neuronal phenotype, to study the appearance of enteric neurons in the developing chicken gut. Immunoreactive cells appear first in the gizzard primordium at E3.5 and are found at progressively more caudal locations in the gut as development proceeds. Nascent neurons are present at the yolk stalk at E4.5, at the ileocecal junction at E6.5, and within the rectum at E7.5-8.5. Neurons appear slightly later in the esophagus. Aggregates of cells resembling developing ganglia were first observed at E6.5 in the distal esophagus and at E8.5 in the proximal esophagus. A small number of cells appeared in the vagus nerve trunks at E4.5 and that number increased at E7.5-8.5. Immunoreactive cells were also found in the sympatho-aortic plexus between the mesonephri and in the dorsal mesentery. These cells appeared to coalesce and form the ganglionated Nerve of Remak which contained positive cells at E3.5. This Nerve extended to the yolk stalk at E4.5 and to duodenum at E6.5. We conclude that the appearance of nascent neurons occurs first in the gizzard and proceeds more rapidly in a distal than proximal direction along the gut. Furthermore, cell that appear to be nascent neurons are found in the vagus and in the dorsal mesentery.
This study was designed to investigate effects of perfusate oxygenation and maturation on coronary flow (CF), interstitial transudate adenosine (ITA) and coronary effluent adenosine (CEA) in isolated rabbit hearts. Hearts were paced at a fixed rate and were perfused under constant pressure at two different levels of perfusate oxygenation: baseline (B) (pO2 = 408 +/- 7 mmHg, O2 content = 1.28 +/- 0.03 ml O2/dl) and a lower level (L) (pO2 = 189 +/- 4 mmHg, O2 content = 0.59 +/- 0.02 ml O2/dl). CF was higher in immature (I, age 5 weeks) compared with mature (M, age 12 weeks) hearts at both levels of perfusate oxygenation (9.7 +/- 0.4 vs. 7.7 +/- 0.3 and 12.9 +/- 0.4 vs. 9.2 +/- 0.3 ml/min/g). I hearts had correspondingly higher values for myocardial oxygen consumption (MVO2) (53 +/- 3 vs. 49 +/- 2 and 39 +/- 3 vs. 35 +/- 2 microL O2/min/g), but similar values for venous oxygen tensions (nuO2) (240 +/- 9 vs. 219 +/- 10 and 74 +/- 4 vs. 62 +/- 3 mmHg), compared with M hearts at B and L. Although interstitial transudate adenosine (ITA) concentration was similar in I and M hearts at B (409 +/- 75 vs. 254 +/- 39 nM), it was lower in I than M hearts at L (2500 +/- 770 vs. 4210 +/- 1000 nM).(ABSTRACT TRUNCATED AT 250 WORDS)
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The enteric nervous system is largely formed from the vagal neural crest which arises from the neuroaxis between somites 1-7. In order to evaluate the contribution of different regions of the vagal crest to the enteric nervous system, we marked crest cells by injecting somites 1-10 with a replication-defective spleen necrosis virus vector which contains the marker gene lacZ. After incubation in X-gal, lacZ-positive blue cells were found in the wall of the gut in three locations. Most were found at the peripheral edge of the developing circular muscle and within the developing submucosa, sites characteristic of developing ganglia. LacZ-positive cells in these ganglionic sites were always surrounded by HNK-1 immunostained cells, confirming their neural crest origin. LacZ-positive cells were also seen in a third location, the circular muscle layer of the esophagus and crop, and were separated from the HNK-1 positive ganglionic elements. These cells in the circular muscle are probably muscle cells derived from labeled mesodermal cells of the somite. Injection of somites 3, 4, 5, and 6 resulted in the largest percentage of preparations with lacZ-positive crest-derived cells and in the largest number of positive cells in the gut. After injection of these somites, lacZ-positive crest-derived cells were found in all regions of the gut from the proventriculus to the rectum. Very few positive crest-derived cells were found in the esophagus.(ABSTRACT TRUNCATED AT 250 WORDS)
In screening a rat pancreatic islet cDNA library by differential hybridization for transcripts increased by glucose, the H chain of ferritin, an iron storage protein, was identified. An ELISA for rat ferritin showed that the insulin cell contains a surprisingly high amount of ferritin comparable to that of iron storage tissues. Most of the ferritin was located in the beta cell, as judged from colocalization of ferritin and insulin by immunohistochemical staining of the pancreas. Islets maintained for 24 h in culture medium containing 20 mM glucose are capable of releasing insulin in response to stimulation with glucose, but islets maintained at 1 mM glucose are incapacitated in response to glucose (see ref 1). Immunoassayable ferritin was threefold higher in 20 mM glucose islets than in 1 mM glucose islets and the glucose-stimulated increase in H ferritin mRNA was confirmed by probing Northern blots of islet RNA with authentic H and L chain cDNAs. This showed that H ferritin mRNA was four- to eightfold higher in 20 mM-glucose islets than in 1 mM glucose islets, whereas L ferritin mRNA was decreased 75-90% in 20 mM glucose islets relative to 1 mM glucose islets. The physiological reason for the abundance of (apo)ferritin in the beta cell is not readily apparent because the islet contains very little iron. Whether or not ferritin is used to handle another metal such as zinc, which is plentiful in the beta cell, is unknown. Another potential reason for ferritin in the beta cell is that ferritin has antioxidant properties and the beta cell is particularly sensitive to oxygen radicals. Regardless of its physiologic purpose, the high amount of ferritin can explain why iron is preferentially retained in the insulin cell and causes diabetes in diseases of iron overloading.
Clinical, echocardiographic, hemodynamic, and angiographic findings were reviewed in 16 consecutive patients with tricuspid atresia (TA) and normally related great arteries, seen at the University of Florida between 1984 and 1990, to determine when and in which hypercyanotic spells (HS), unrelated to ductal constriction, occurred. Such spells developed in six patients (38%) and were life-threatening and refractory to medical management in three patients. Most HS (five of six) occurred in the first 2 months of life. Angiography done at the time of diagnosis revealed similar morphological features in all patients subsequently experiencing HS; infundibular stenosis due to anterior malalignment in the conal septum. These findings suggest a predisposition for development of early HS in TA patients with apparent infundibular stenosis due to anterior malalignment of the conal septum. Further study and a heightened clinical awareness are needed in such patients.