Plant lignans inhibit growth and trichothecene biosynthesis in Fusarium graminearum.

UNLABELLED: Lignans are a group of diphenolic compounds with anticancer and antioxidant properties which are present in various grains, although their effect on toxigenic fungi has been poorly examined to date. In this study, the impact of the plant lignans pinoresinol and secoisolariciresinol on growth and trichothecene biosynthesis by five Fusarium graminearum strains of different chemotypes was examined in vitro. Both tested lignans exhibited radial growth inhibition against the fungal strains. RT-qPCR analyses of tri4, tri5 and tri11 genes encoding the first steps of the trichothecene biosynthesis pathway revealed a decrease in tri mRNA levels in lignan-treated fungal cultures. Correspondingly, decreased accumulation of toxins in lignan-treated cultures was confirmed by GC-MS analysis. This is the first study to demonstrate the inhibitory effect of both pinoresinol and secoisolariciresinol on growth and trichothecene biosynthesis in F. graminearum. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of the regulation of trichothecene production in Fusarium graminearum by environmental cues is key to the design of novel strategies to reduce mycotoxin levels in grains. Here, we show that the lignans pinoresinol and secoisolariciresinol, which occur in wheat grains, inhibit radial growth and decrease trichothecene levels in five F. graminearum strains. RT-qPCR analysis reveals that the reduction in trichothecene level in lignan-treated fungal cultures is associated with decreased mRNA transcript levels for the tri4, tri5 and tri11 genes that are involved in the trichothecene biosynthesis pathway.

Predictors of quality of life in peri- and postmenopausal Polish women living in Lublin Voivodeship.

OBJECTIVES: The aim of this observational cross-sectional study was to establish the factors that determine the quality of life in a sample of peri- and postmenopausal women and to answer the question of whether the quality of life of these women is dependent on currently or previously received hormone replacement therapy (HRT). METHODS: The research was carried out by means of a survey method, postal questionnaire technique. Three standardized questionnaires: WHOQOL-BREF, Women's Health Questionnaire (WHQ) and SF-36 were used as research tools. An original questionnaire was also used. The study comprised a representative sample of the female population aged 45-65 years living in Lublin Province. The sample size was 2143 women. The domains of quality of life established by the WHOQOL-BREF, WHQ and SF-36 questionnaires were treated as dependent variables, whereas the sociodemographic variables, data concerning the women's gynecological history, their state of health and whether they received HRT or not were treated as independent variables. RESULTS: At multivariate analysis, self-assessment of the state of health as poor or fair, the presence of urinary incontinence, the presence of chronic diseases, self-assessment of living conditions as poor, self-assessment of financial situation as poor, eligibility for benefits (pensions) for the disabled, and lower education level represented the most important predictors of poor quality of life. HRT use had an independent impact on women's quality of life only in one quality-of-life domain - sleep problems in the WHQ. Current HRT users were characterized by a slightly lower risk of quality of life reduction when compared with past HRT users and women who never used HRT. CONCLUSIONS: Strong predictors of the worse quality of life established in the research make it possible to single out a group of women who need special attention in the process of undertaking preventive or curative steps.

The evaluation of the efficacy of sodium carbonate as zearalenone destructor in feeding stuffs.

Contamination of feed with zearalenone (ZEA) is still a serious problem in farm animals feeding, especially in gilts, sensitive to this compound. The relative failure of current methods of decontamination and quality control lead us to look for new techniques. The commonly accepted method for breaking down ZEA was performed in controlled temperature and time conditions. Various sodium carbonate doses (0.5 - 4%) were added to feed naturally contaminated with ZEA (ZEA biosynthesis by F. graminearum isolates). These doses were found to be effective in in vitro studies. The addition of 2% sodium carbonate gave the best results in reducing the phytoestrogen in the feed.

Structural features of polymer adsorbent LiChrolut EN and interfacial behavior of water and water/organic mixtures.

The structural and adsorption characteristics of polymer adsorbent LiChrolut EN and the behavior of adsorbed water and water/organic mixtures were studied using adsorption, microcalorimetry, transmission and scanning electron microscopy, mass spectrometry, infrared spectroscopy, 1H NMR spectroscopy with layer-by-layer freezing-out of liquids (190-273 K), and thermally stimulated depolarization current method (90-265 K). This adsorbent is characterized by large specific surface area (approximately 1500 m2/g) and pore volume (0.83 cm3/g) with a major contribution of narrow pores (R<10 nm) of a complicated shape (long hysteresis loop is in nitrogen adsorption-desorption isotherm). The adsorbent includes aromatic and aliphatic structures and oxygen-containing functionalities and can effectively adsorb organics and water/organic mixtures. On co-adsorption of water and organics (dimethyl sulfoxide, chloroform, methane), there is a weak influence of one on another adsorbate due to their poor mixing in pores. Weakly polar chloroform displaces a fraction of water from narrow pores. These effects can explain high efficiency of the adsorbent in solid-phase extraction of organics from aqueous solutions. The influence of structural features of several carbon and polymer adsorbents on adsorbed nitrogen, water and water/organics is compared on the basis of the adsorption and 1H NMR data.

Close similarity among streptavidin-like, biotin-binding proteins from Streptomyces.

Two strains of Streptomyces venezuelae were found to produce high-affinity, biotin-binding proteins, termed streptavidin v1 and v2, respectively. Both proteins were isolated to purity, and their corresponding genes were cloned and sequenced. Compared to streptavidin from S. avidinii, streptavidin v1 had only a single amino acid substitution and streptavidin v2 showed 9 such differences. The substitutions were remarkably conservative, none of which affected the amino acid residues known to be important to the biotin-binding properties or to the structure of the tetrameric protein. The results also indicate that the biosynthesis of such biotin-binding proteins is not simply a curious anomaly in a single species of Streptomyces. It is suggested that the classification of S. avidinii as a unique species should be reconsidered. The occurrence of these proteins appears to be linked to the production of an unusual synergistic antibiotic complex.

Balloon dilation of the aortic valve: studies in normal lambs and in children with aortic stenosis.

To evaluate the risks of and optimal method for valve dilation in aortic stenosis, balloons of different sizes were used to dilate the normal aortic root in 16 lambs and then stenotic valves in 15 children. In the lambs, inflated balloon to aortic anulus diameter ratios ranged from 0.9 to 1.5. These hearts were examined immediately after the procedure. Ratios of 0.9 to 1.1 did not produce significant damage to the left ventricular outflow tract, whereas those of 1.2 to 1.5 produced tears or hematomas, or both, of the aortic valve leaflets (n = 3), mitral valve leaflets (n = 4) and interventricular septum (n = 4). The 15 patients, aged 10 days to 15 years, underwent 16 balloon aortic valvotomy procedures. The balloon-aortic anulus ratio ranged from 0.67 to 1.1 (mean 0.90). The average pressure gradient decreased 69% and, overall, the peak systolic gradient decreased from 86 +/- 21 to 28 +/- 14 mm Hg (p less than 0.01) and the aortic valve area increased from 0.44 +/- 0.11 to 0.73 +/- 0.22 cm2/m2 (p less than 0.01). Immediately after the procedure an increase in aortic regurgitation was noted in 8 (57%) of 14 patients, but was never greater than 3+ and has been well tolerated. Other early complications encountered consisted of transient left bundle branch block in two patients, temporary femoral artery occlusion in three and femoral artery rupture requiring operative management in one infant. Balloon valvotomy can reduce the transvalvular gradient in most patients with valvular aortic stenosis when a balloon less than 1.1 times the aortic root diameter is used.(ABSTRACT TRUNCATED AT 250 WORDS)

Morphology and surface properties of fumed silicas.

Several series of fumed silicas and mixed fumed oxides produced and treated under different conditions were studied in gaseous and liquid media using nitrogen and water adsorption-desorption, mass spectrometry, FTIR, NMR, thermally stimulated depolarization current (TSDC), photon correlation spectroscopy (PCS), zeta potential, potentiometric titration, and Auger electron spectroscopy methods. Aggregation of primary particles and adsorption capacity (Vp) decrease and hysteresis loops of nitrogen adsorption-desorption isotherms becomes shorter with decreasing specific surface area (S(BET)). However, the shape of nitrogen adsorption-desorption isotherms can be assigned to the same type independent of S(BET) value. The main maximum of pore size distribution (gaps between primary nonporous particles in aggregates and agglomerates) shifts toward larger pore size and its intensity decreases with decreasing S(BET) value. The water adsorption increases with increasing S(BET) value; however, the opposite effect is observed for the content of surface hydroxyls (in mmol/m2). Associative desorption of water (2(SiOH)-->SiOSi+H2O) depends on both the morphology and synthesis conditions of fumed silica. The silica dissolution rate increases with increasing S(BET) and pH values. However, surface charge density and the modulus of zeta-potential increase with decreasing S(BET) value. The PCS, 1H NMR, and TSDC spectra demonstrate rearrangement of the fumed silica dispersion depending on the S(BET) value and the silica concentration (C(SiO2)) in the aqueous suspensions. A specific state of the dispersion is observed at the C(SiO2) values corresponding to the bulk density of the initial silica powder.

Recombinant NeutraLite avidin: a non-glycosylated, acidic mutant of chicken avidin that exhibits high affinity for biotin and low non-specific binding properties.

A recombinant non-glycosylated and acidic form of avidin was designed and expressed in soluble form in baculovirus-infected insect cells. The mutations were based on the same principles that guided the design of the chemically and enzymatically modified avidin derivative, known as NeutraLite Avidin. In this novel recombinant avidin derivative, five out of the eight arginine residues were replaced with neutral amino acids, and two of the lysine residues were replaced by glutamic acid. In addition, the carbohydrate-bearing asparagine-17 residue was altered to an isoleucine, according to the known sequences of avidin-related genes. The resultant mutant protein, termed recombinant NeutraLite Avidin, exhibited superior properties compared to those of avidin, streptavidin and the conventional NeutraLite Avidin, prepared by chemo-enzymatic means. In this context, the recombinant mutant is a single molecular species, which possesses strong biotin-binding characteristics. Due to its acidic pI, it is relatively free from non-specific binding to DNA and cells. The recombinant NeutraLite Avidin retains seven lysines per subunit, which are available for further conjugation and derivatization.

Mutation of a critical tryptophan to lysine in avidin or streptavidin may explain why sea urchin fibropellin adopts an avidin-like domain.

Sea urchin fibropellins are epidermal growth factor homologues that harbor a C-terminal domain, similar in sequence to hen egg-white avidin and bacterial streptavidin. The fibropellin sequence was used as a conceptual template for mutation of designated conserved tryptophan residues in the biotin-binding sites of the tetrameric proteins, avidin and streptavidin. Three different mutations of avidin, Trp-110-Lys, Trp-70-Arg and the double mutant, were expressed in a baculovirus-infected insect cell system. A mutant of streptavidin, Trp-120-Lys, was similarly expressed. The homologous tryptophan to lysine (W-->K) mutations of avidin and streptavidin were both capable of binding biotin and biotinylated material. Their affinity for the vitamin was, however, significantly reduced: from K(d) approximately 10(-15) M of the wild-type tetramer down to K(d) approximately 10(-8) M for both W-->K mutants. In fact, their binding to immobilized biotin matrices could be reversed by the presence of free biotin. The Trp-70-Arg mutant of avidin bound biotin very poorly and the double mutant (which emulates the fibropellin domain) failed to bind biotin at all. Using a gel filtration fast-protein liquid chromatography assay, both W-->K mutants were found to form stable dimers in solution. These findings may indicate that mimicry in the nature of the avidin sequence and fold by the fibropellins is not designed to generate biotin-binding, but may serve to secure an appropriate structure for facilitating dimerization.

Engineering of chicken avidin: a progressive series of reduced charge mutants.

Avidin, a positively charged egg-white glycoprotein, is a widely used tool in biotechnological applications because of its ability to bind biotin strongly. The high pI of avidin (approximately 10.5), however, is a hindrance in certain applications due to non-specific (charge-related) binding. Here we report a construction of a series of avidin charge mutants with pIs ranging from 9.4 to 4.7. Rational design of the avidin mutants was based on known crystallographic data together with comparative sequence alignment of avidin, streptavidin and a set of avidin-related genes which occur in the chicken genome. All charge mutants retained the ability to bind biotin tightly according to optical biosensor interaction analysis. In most cases, their thermal stability characteristics were indistinguishable from those of the wild-type avidin. Our results demonstrate that the charge properties of avidin can be modified without disturbing the crucial biotin-binding activity.

Morphologic changes induced by dilation of the pulmonary valve anulus with overlarge balloons in normal newborn lambs.

The optimal method for balloon dilation valvuloplasty of congenital pulmonary valve stenosis has not been established. The cardiac damage produced by using a balloon the same size as the pulmonary anulus has not been described. The use of balloons that are larger than the anulus or of a dynamic dilation technique may enhance gradient reduction, but their added risks are not known. The pulmonary valve anuli of 14 normal newborn lambs were dilated with angioplasty balloons 20% smaller to 90% larger than the anulus; in 3 dilations, a dynamic technique was used, consisting of withdrawal of the fully inflated balloon from the anulus into the body of the right ventricle. Twelve lambs were killed immediately after the procedure and 2 were killed later, and detailed gross anatomic and microscopic observations of the heart were made. These observations support the following conclusions: (1) Dilation of the pulmonary valve anulus with overlarge balloons is "clinically" well tolerated in normal newborn lambs. (2) The major damage to the heart from any balloon is not the pulmonary anulus, but to the right ventricular outflow tract and free wall, with mural hemorrhages of varying size. (3) Trauma is minor if the balloon is 30% larger than the anulus, but is considerable when the balloon is 50% larger; (4) Trauma is worst subjacent to the proximal end of the balloon, so that longer balloons may cause greater damage; and (5) Resolution of acute right ventricular hemorrhage occurs with small, patchy areas of fibrosis.

The "laid-back" aortogram: an improved angiographic view for demonstration of coronary arteries in transposition of the great arteries.

Preoperative demonstration of coronary arterial anatomy may be important for babies undergoing the arterial switch operation. Echocardiography is clearly useful, but may not unequivocally show all coronary branches. Standard angiographic views can be confusing. An improved angiographic projection in which the frontal x-ray tube is caudally angled, resulting in a "laid-back" position of the image intensifier and cine camera, provides superior visualization of the coronary arteries and their relation to the aorta and the pulmonary artery. The balloon occlusion technique is used for opacification of the aortic root from the transvenous approach. Injection of 1 ml/kg of contrast delivered in 1/2 to 1 second provides the best images. The caudal aortogram is easier to interpret than standard views and facilitates description and recognition of various coronary patterns. The relation between the pulmonary artery and the aorta, the origins of the coronary arteries from the facing sinuses and their proximity to the intercoronary commissures, and the myocardial distribution of each coronary vessel are shown clearly. The caudal view therefore offers significant advantages over conventional projections for demonstration of coronary arterial anatomy in infants with transposition of the great arteries or double-outlet right ventricle.

Chronic percutaneous pericardial drainage with modified pigtail catheters in children.

To determine the safety and efficacy of chronic percutaneous pericardial drainage in children, pigtail catheters were inserted over curved guidewires under fluoroscopic control into the pericardial space in 7 consecutive children with pericardial effusion. Pericardiocentesis was therapeutic (for tamponade) in 1 child, diagnostic in 4 and both therapeutic and diagnostic in 2. The children were 0.5 to 16 years old and weighed 5 to 65 kg. Underlying diagnoses included cancer (3 children), congenital heart disease (2 children) and immunodeficiency and hemolytic uremic syndrome (1 each). When unmodified pigtail catheters, designed for angiography, were used (as in the first 3 children), either the catheters clotted within 36 hours, necessitating operative pericardial drainage, or repeated heparin infusions were required to keep the catheter patent. However, when 8Fr catheters were modified by placing 0.050-inch side holes along the distal shaft, the catheters remained patent and effectively drained the pericardial space for 3 to 7 days. Heparin infusion was not required, no child managed with the modified catheters required subsequent drainage and no complications occurred. In conclusion, percutaneous pericardial drainage is safe, even in small children, and can be effective chronically if catheters with large drainage holes are used.

Postoperative hemodynamics after Norwood palliation for hypoplastic left heart syndrome.

Hemodynamics after Norwood palliation for hypoplastic left heart syndrome (HLHS) have been incompletely characterized, although emphasis has been placed on the role that an excess pulmonary-to-systemic blood flow ratio (Qp/Qs) may play in causing hemodynamic instability. Studies suggest that maximal oxygen delivery occurs at a Qp/Qs < 1. However, it remains unclear to what extent cardiac output can increase with increasing pulmonary perfusion. One approach is to use the oxygen excess factor omega, an index of systemic oxygen delivery, and compare omega with measured Qp/Qs. We measured Qp/Qs and omega in neonates after Norwood palliation for HLHS, and determined how they were related. In addition, we determined the temporal course of surrogate indexes of systemic perfusion in the early postoperative period. Arteriovenous oxygen saturation difference, blood lactate, and omega were recorded on admission and every 3 to 12 hours for 2 days in 18 consecutive infants with HLHS or variant after Norwood palliation. Three infants required extracorporeal membrane oxygenation (ECMO) 6 to 9 hours after admission. These infants had higher Qp/Qs, blood lactate, arteriovenous oxygen saturation difference, and lower omega than non-ECMO patients. In non-ECMO patients between admission and 6 hours, omega decreased significantly despite no appreciable change in Qp/Qs. We conclude that: (1) Oxygen delivery is significantly decreased at 6 postoperative hours unrelated to Qp/Qs. This modest decline in oxygen delivery is insufficient to compromise tissue oxygenation. (2) Patients requiring ECMO have significant derangements in oxygen delivery.

Collagen content in normal, pressure, and pressure-volume overloaded developing human hearts.

Increased myocardial collagen accompanies pressure overload of the adult left ventricle. This phenomenon is poorly understood in infants. This study compares the myocardial volume fraction of collagen in infants who did not have primary heart disease with infants with isolated pressure overload of the right ventricle (tetralogy of Fallot [ToF]), and with infants with combined volume and pressure overload (aortic valve atresia [AVA]). The distribution of collagen in the neonatal myocardium was also determined. We measured the volume fraction of collagen from right ventricular biopsy specimens of cadaver hearts in normal infants (1 to 9 months old; n = 7), infants with ToF (1 day to 9 months old; n = 9), newborns with AVA (AVA-NB) (1 to 4 days old; n = 5), and older patients with AVA (AVA-I) (5 to 8 months old; n = 5). Myocardium from 3 patients undergoing repair of ToF (6 to 8 months old) was also analyzed. Specimens were stained with Masson's trichrome and myocardial volume fraction of collagen determined by point counting. Myocardial volume fraction of collagen was significantly higher (p = 0.02) in AVA-I patients (8.0 +/- 3.5%) versus normal (3.3 +/- 2.7%), ToF (3.2 +/- 1.8%), and AVA-NB (3.5 +/- 2.3%) patients. There was a tendency for increased collagen in the subendocardium, especially in AVA-I patients (p > 0.05). We conclude that patients with AVA-I have increased collagen relative to normal subjects, patients with ToF, and patients with AVA-NB, and that this increase is greatest in the subendocardium.

Cardiac arrest in young, ostensibly healthy patients: clinical, hemodynamic, and electrophysiologic findings.

This study examines the clinical, hemodynamic, and electrophysiologic findings in a unique group of 11 young (aged 15 months to 29 years) survivors of a cardiac arrest. All patients were previously in good health, and cardiac arrest was the initial manifestation of cardiac disease in all. Overt clinical and hemodynamic abnormalities were not as common as previously reported, and in some instances apparent cardiac abnormalities failed to provide a link to cardiac arrest. No patient had congenital heart disease or hypertrophic cardiomyopathy. However, during multicatheter electrophysiologic study, sustained tachyarrhythmia was reproducibly initiated in 8 of 11 patients (73%). Young, ostensibly healthy patients who survive cardiac arrest form a diverse group. Diligent programmed intracardiac electrical stimulation may demonstrate life-threatening tachycardias in these patients. Treatment to prevent recurrence of cardiac arrest is difficult in this group of patients. However, the ability to initiate tachycardia in the electrophysiologic laboratory may be useful in the management of these patients.

Carboxy derivatives of isoflavones as affinity carriers for cytotoxic drug targeting in adrenocortical H295R carcinoma cells.

Carboxy derivatives of isoflavones that exhibit oestrogenic/anti-oestrogenic properties were used as carriers for affinity drug targeting to H295R adrenocortical carcinoma cells that express transcripts of oestrogen receptor (ER) alpha and beta. These derivatives were prepared by introducing a carboxymethyl group at the 6-position of genistein and of biochanin A, yielding 6CG and 6CB respectively. In transactivation assays, 6CG displayed mixed agonist/antagonist activity for ERalpha, whereas 6CB displayed only weak antagonist activity. Low concentrations of oestrogen, 6CG and 6CB were capable of inducing proliferation in H295R cells and of stimulating creatine kinase (CK) specific activity, suggesting that these cells were sensitive to oestrogenic compounds. In in vivo experiments, both 6CG and 6CB were capable of inhibiting oestrogen-induced CK specific activity in rat tIssues. For affinity drug targeting, the cytotoxic drug daunomycin was coupled to 6CB and 6CG, yielding 6CB-Dau and 6CG-Dau respectively. These conjugates were tested for their antiproliferative ability to inhibit DNA synthesis as assessed by incorporation of [(3)H]thymidine in H295R cells. A dose-dependent cytoxicity was observed with both conjugates. At 0.3-3 nM, both conjugates were 10 to 30 times more potent than daunomycin. At 30 nM these conjugates were two to three times more potent than daunomycin. At concentrations ranging between 300 and 3000 nM, no difference in cytotoxicity was observed between the conjugates and daunomycin. When the cells were treated over a wide range of concentrations with a combination of 6CG plus daunomycin, the observed cytotoxicity was less than with daunomycin alone. When non-transformed rat enterocytes, which do not express ER, were treated with 6CG-Dau or daunomycin, the antiproliferative effect of 6CG-Dau was the same as that of daunomycin over the concentration range tested. These pilot studies suggest that the ready availability of oestrogenic binding sites in H295R cells can be exploited for site-directed chemotherapy.

Role of putative membrane receptors in the effect of androgens on human vascular cell growth.

We have reported previously that dihydrotestosterone (DHT) induces a biphasic effect on DNA synthesis in human vascular smooth muscle cells (VSMC), i.e. stimulation at low concentrations and inhibition at high concentrations. In contrast, DHT dose-dependently stimulated [(3)H]thymidine incorporation in a human endothelial cell line (ECV304). Additionally, DHT increased the specific activity of creatine kinase (CK) in both vascular cell types. In the present study, we have determined whether some of these effects are exerted via membrane-binding sites. We measured changes in DNA synthesis and CK after treatment with DHT and the membrane-impermeant testosterone-3-carboxymethyl oxime conjugated to bovine serum albumin (BSA) (T-BSA). High concentrations of either DHT or T-BSA inhibited VSMC proliferation (by 52+22% and 51+25% respectively). DHT as well as T-BSA increased DNA synthesis in ECV304 cells dose-dependently. In contrast, T-BSA did not affect CK in either cell type. In both cell types, DHT as well as T-BSA increased mitogen-activated protein kinase (MAPK) kinase activity as measured by total phosphorylated MAPK. Further, the inhibitory effect of either the free or protein-bound androgens on DNA synthesis was blocked by UO126, an inhibitor of MAPK kinase activity. T-BSA conjugate labeled with Europium showed binding to whole VSMC, which could be displaced by excess T-BSA, but not by estradiol-BSA or the free hormones. Finally, using T-BSA linked to the fluorescent dye Cy3.5, we directly demonstrated the presence of membrane-binding sites for androgen in VSMC. Hence, the inhibitory effects of testosterone on DNA synthesis in VSMC are apparently exerted by membrane-binding sites for androgen, do not require intracellular entry of the hormone and its binding to the classical nuclear receptors and are linked to MAPK activation.

6-Carboxymethyl genistein: a novel selective oestrogen receptor modulator (SERM) with unique, differential effects on the vasculature, bone and uterus.

The novel genistein (G) derivative, 6-carboxymethyl genistein (CG) was evaluated for its biological properties in comparison with G. Both compounds showed oestrogenic activity in vitro and in vivo. On the other hand G and CG differed in the following parameters: (i) only CG displayed mixed agonist-antagonist activity for oestrogen receptor (ER) alpha in transactivation assays and (ii) only CG was capable of attenuating oestrogen (E(2))-induced proliferation in vascular smooth muscle cells and of inhibiting oestrogen-induced creatine kinase (CK) specific activity in rat tissues. On the other hand only G enhanced the stimulatory effect on CK specific activity in the uterus. In comparison to the selective oestrogen receptor modulator (SERM) raloxifene (RAL), CG showed the same selectivity profile as RAL in blocking the CK response to E(2) in tissues derived from both immature and ovariectomized female rats. Molecular modelling of CG bound to the ligand binding domain (LBD) of ERbeta predicts that the 6-carboxymethyl group of CG almost fits the binding cavity. On the other hand, molecular modelling of CG bound to the LBD of ERalpha suggests that the carboxyl group of CG may perturb the end of Helix 11, eliciting a severe backbone change for Leu 525, and consequently induces a conformational change which could position Helix 12 in an antagonist conformation. This model supports the experimental findings that CG can act as a mixed agonist-antagonist when E(2) is bound to its receptors. Collectively, our findings suggest that CG can be considered a novel SERM with unique effects on the vasculature, bone and uterus.

Correction of total anomalous pulmonary venous connection in infancy.

From January 1985 through January 1993, 41 patients less than 1 year of age underwent operative correction of isolated total anomalous pulmonary venous connection. There were 24 boys and 17 girls. The median age at operation was 13 days (range 1 to 282 days) and weight was 3.6 kg (2.5 to 5.2 kg). Locations of the connections were supracardiac in 19, cardiac in 9, infracardiac in 11, and mixed supracardiac and cardiac in 2. Obstruction of the pulmonary veins was severe in 24, mild in 3, and absent in 14. Preoperative stabilization included mechanical ventilation for 15 patients for a mean duration of 2 1/2 days and extracorporeal membrane oxygenation for 1 patient for 1 day. All operations were performed with deep hypothermia and circulatory arrest (mean arrest time 34 minutes). Supracardiac connections were repaired by performing a side-to-side anastomosis between the pulmonary venous confluence and the dome of the left atrium through a superior approach between the superior vena cava and the aorta. Coronary sinus connections were repaired by enlarging the atrial septal defect and the coronary sinus communication with the left atrium and closing the atrial defect with a large patch. Infracardiac repairs included elevation and rotation of the heart to the right and an elongated side-to-side anastomosis between the common venous confluence and the left atrium. One patient died 1 week postoperatively of persistent pulmonary hypertension. Another patient, who was supported by extracorporeal membrane oxygenation before the operation, died 3 months after the operation as a consequence of pulmonary lymphangiectasia. All other patients are alive and well with a mean follow-up of 26 months (range 3 to 77 months). One patient required two subsequent reoperations for persistent pulmonary venous obstruction, and another patient had superior vena cava obstruction necessitating reoperation. Operative treatment of total anomalous pulmonary venous connection in infants can be performed with low mortality and an infrequent need for reoperations.