A novel protein LUZP with 3 leucine zipper motifs at its amino terminus is predominantly expressed in the adult brain. A modified gene targeting approach was employed in an attempt to establish in vitro and in vivo models in which Luzp is knock-out (KO) for phenotype assessment and a reporter gene lacZ is knock-in (KI) for tracing its expression. We report in this study the molecular cloning of the Luzp gene, its targeting vector construction and Luzp-KO/lacZ-KI embryonic stem (ES) clone selection. Since LUZP is also expressed in ES cells, the possibility of embryonic lethality cannot be excluded when attempting to establish Luzp-null mutant mice. We have therefore examined the development of homozygous Luzp-KO/lacZ-KI clones in nude mice. Tissue types derived from all three embryonic germ layers were observed in teratomas developed in nude mice. In situ X-gal staining further revealed restricted expression of LUZP in neural lineage cells.

Animals↗

PubMedDate not supplied

Mechanical cardiopulmonary support in children and young adults: extracorporeal membrane oxygenation, ventricular assist devices, and long-term support devices.

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Adolescent↗

PubMedDate not supplied

Left ventricular noncompaction cardiomyopathy in association with trisomy 13.

In recent years, left ventricular noncompaction (LVNC) has been recognized as a distinct form of cardiomyopathy with its own clinical presentation and natural history. More than 100 cases of LVNC have been described in children. Although LVNC has been described in association with metabolic disorders such as Fabry's disease or genetic disorders such as Roifman's syndrome, this case represents the first report of LVNC in a child with trisomy 13.

Cardiomyopathy, Dilated↗

PubMedDate not supplied

Captopril-induced pulmonary infiltrates with eosinophilia in an infant with congenital heart disease.

We report a case of an infant with complex congenital heart disease who was placed on captopril for afterload reduction following cardiac surgery and subsequently developed pulmonary infiltrates with eosinophilia. The patient was readmitted with symptoms of rhinorrhea, poor feeding, and decreased activity level. She was found to have diffuse pulmonary infiltrates on chest radiograph and a marked peripheral eosinophilia without leukocytosis. After discontinuing captopril and starting systemic steroids, her symptomatology rapidly improved, and her eosinophilia and radiographic abnormalities both resolved.

Angiotensin-Converting Enzyme Inhibitors↗

PubMedDate not supplied

A prospective evaluation of nesiritide in the treatment of pediatric heart failure.

This study sought to determine the potential of recombinant B-type natriuretic peptide (nesiritide) for the treatment of pediatric decompensated heart failure. Nesiritide is a widely used and effective treatment for decompensated heart failure (HF) in adults, but its safety and efficacy in pediatric patients is unclear. Outcomes of 55 separate nesiritide infusions of varying durations in 32 patients (13 males and 19 females; mean age, 8.01 years; range, 0.01-20.4) were evaluated prospectively. All patients received nesiritide in the intensive care unit. The starting dose (0.01 microg/kg/min) was titrated to a maximum of 0.03 microg/kg/min. All patients were monitored for clinical signs and symptoms, hemodynamics, urine output, electrolytes, oxygen requirements, and oral intake. Functional status was assessed by patients and/or their parents. All patients successfully underwent initiation and titration of nesiritide infusion. No hypotension or arrhythmias were noted during 478 cumulative days of therapy. Nesiritide was given safely with vasoactive medications. Mean urine output improved from 2.35 +/- 1.71 cc/kg/hr on the day before nesiritide initiation (baseline) to 3.10 +/- 1.94 cc/kg/hr on day 4 of treatment (p < 0.01). Serum creatinine decreased from 1.04 to 0.92 mg/dl (p = 0.096), mean central venous pressure from 13 to 7 mmHg (p = 0.018), and mean weight from 30.4 to 29.7 kg (p < 0.001) with therapy. Thirst, as subjectively assessed by patients old enough to respond, decreased with infusion in 31 of 42 cases (74%). Mean New York Heart Association functional class improved significantly (p < 0.001). Nesiritide infusion, alone or in combination, is a safe treatment for decompensated HF in pediatric patients. It is associated with decreased thirst and improved urine output and functional status, and it may be efficacious in the treatment of pediatric HF.

Adolescent↗

PubMedDate not supplied

Adverse hemodynamic effects observed with inhaled nitric oxide after surgical repair of total anomalous pulmonary venous return.

The following is a case report of a 1-month-old patient who developed adverse hemodynamic sequelae during the use of nitric oxide (NO) in the postoperative period for pulmonary hypertension after correction of total anomalous pulmonary venous return. At the time of diagnosis, the patient had evidence of systemic right ventricular pressures estimated by continuous-wave Doppler. He was sedated and paralyzed for hyperventilation in preparation for surgery and underwent pulmonary vein confluence to left atrial anastomosis. Postoperative pulmonary hypertension was managed by hyperventilation, sedation, and paralysis until a sudden onset of systemic-level pulmonary pressure required NO therapy. Satisfactory results were obtained in minutes, but a rebound pulmonary hypertension occurred with concomitant systemic hypertension and no radiographic changes. We suspected left atrial hypertension secondary to a sudden increase in pulmonary blood flow to an noncompliant left ventricle. Discontinuation of NO resulted in stabilization of the hemodynamic profile of the patient and he continued to be managed with paralysis, hyperventilation, and sedation. Based on this experience we suggest that NO should be used with caution in patients with obstructive lesions at the atrial level prior to surgery (mitral valve stenosis and cor triatriatum) or in patients with a poorly compliant left ventricle (cardiomyopathy and left ventricular dysfunction). These entities are unable to tolerate a sudden increase in pulmonary blood return thus creating paradoxical pulmonary hypertension.

Administration, Inhalation↗

PubMedDate not supplied

Prolonged anti-CD40 ligand therapy improves primate cardiac allograft survival.

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Animals↗

PubMedDate not supplied

Determination of protein loading in biodegradable polymer microspheres containing tetanus toxoid.

Various methods to determine loading of vaccine in biodegradable polymer microspheres encapsulating tetanus toxoid were evaluated. The microspheres were composed of poly (D-lactic acid) (PLA) and poly (DL-lactic-co-glycolic acid) (PLGA). Dissolution of microspheres in organic solvents such as methylene chloride, chloroform, or dimethyl sulfoxide and extraction of vaccine antigen or total protein with phosphate buffered saline gave variable results which depended upon the characteristics of the microspheres, such as type of polymer, excipients used in the microspheres and formulation conditions. Microspheres made from low molecular weight PLGA polymer and showing a large burst release exhibited up to 25% extraction of antigen whereas microspheres made from PLA microspheres with low burst release showed < 1% extraction. Extraction of total protein with 0.1 N NaOH and 5% sodium dodecyl sulfate showed results similar to those obtained with organic solvent extraction method. Partial digestion of microspheres with 6 N HCl at 60 degrees C for 20 h resulted in approximately 30% loss in TT protein by micro-bicinchoninic acid (BCA) assay. The major problem with this method was strong reactions in the micro-BCA assay of stabilizers, particularly sugars (glucose, sucrose) used in the microsphere formulations. Complete digestion of microspheres with 6 N HCl at 110 degrees C for 20 h or with 13.5 N NaOH at 121 degrees C for 1 h and quantitation of amino acids by a modified ninhydrin assay showed reproducible results on the protein loading in the microspheres. However, this method was affected by the presence of stabilizers, such as gelatin, which contain amino acids. Further, sucrose concentrations higher than 10% caused interference in the ninhydrin assay on samples hydrolyzed with 6 N HCl. In contrast, hydrolysis with 13.5 N NaOH did not show any interference by sucrose. Stabilizers used outside the microspheres for lyophilization purposes may be removed by washing the microspheres before loading determination or by dialysis but stabilizers used inside the microspheres would still cause interference. For reliable determination of total protein in the microspheres containing vaccines, we suggest complete digestion of microspheres with acid or base followed by amino acid analysis by colorimeteric assays such as ninhydrin method or using amino acid analyzers. The method needs to be optimized for each type of formulation to eliminate interference by the excipients. Alternatively, total protein nitrogen in the microspheres may be determined by the Kjel-dahl method if no amino acids or other nitrogen containing stabilizer is used inside the microspheres.

Antigens, Bacterial↗

PubMedDate not supplied

Comparison of rejection in the atrioventricular node and bundles with the working myocardium in transplanted hearts.

At present the monitoring of heart transplant patients for rejection is done by endomyocardial biopsy. It has been proposed that ECG assessments of conduction delay may provide a noninvasive means of detecting rejection. To determine whether rejection in the atrioventricular node and conduction bundles reflects rejection in the working myocardium, we studied 21 transplanted hearts from 21 cardiac allograft recipients. Twenty of the hearts were obtained at autopsy, and one was obtained from a patient who underwent a second transplant procedure. The atrioventricular conduction tissues, the ventricular and atrial free walls, and the interventricular and interatrial septa from these hearts were examined for histologic evidence of rejection. Of the 21 hearts, 17 showed histologic changes of acute allograft rejection. Of the 17, rejection involved the conducting system and the myocardium equally in 11. In 6 of the 17 hearts, rejection involved the working myocardium more severely than it did the conducting system. Of interest, nonrejection pathologic changes were also noted in the conducting systems of several hearts. Severe accelerated arteriosclerosis was found in the artery to the atrioventricular node in one case, and lesser degrees of accelerated arteriosclerosis in this artery were found in two additional cases. In one case, lymphoid infiltrates, consistent with posttransplant lymphoproliferative disorder, were noted in the vicinity of the atrioventricular node, and several hearts demonstrated operative trauma or ischemic changes that appeared to involve the conducting system. These results suggest that although there may be a morphologic basis for using electrophysiologic changes in the conducting system to monitor heart allograft recipients for rejection, caution should be exercised in interpreting these changes.

Adult↗