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Biomedical subjects

R Lee

Publications and source records attributed to R Lee.

At least 199 records · Page 11Linked to original sources

Cystic fibrosis heterozygote screening in 5,161 pregnant women.

A screening program for cystic fibrosis (CF) heterozygotes was conducted in a large HMO prenatal population, to evaluate the level of interest among eligible patients, the effectiveness of prescreening education, attitudes toward the screening process, psychological effects, and utilization of prenatal diagnosis and its outcomes. The heterozygote identification rate and frequency of specific CFTR mutations were also assessed. Identified carriers were offered genetic counseling and testing of male partners. Prenatal diagnosis was offered if both parents were identified as carriers. A total of 5,161 women underwent carrier testing; 947 others completed survey instruments only. The acceptance rate of screening was high (78%), and pretest education by videotape was generally effective. Adverse psychological effects were not reported. Participants generally found screening to be desirable and useful. Screening identified 142 female heterozygotes, 109 couples in which the male partner was not a carrier, and 7 high-risk couples. The incidence of R117H mutations was much higher than expected. The number of identified carriers was much lower in Hispanics than in Caucasians. We conclude that large-scale prenatal screening for CF heterozygotes in the absence of a family history of CF is an acceptable method for identifying couples at risk for affected fetuses. Sufficient pretest education can be accomplished efficiently, test insensitivity is well accepted, adverse psychological events are not observed, and general patient satisfaction is high.

Adult↗

UHG-based mutation screening in type 2B von Willebrand's disease: detection of a candidate mutation Ser547Phe.

We have recently described a novel mutation screening technique for the diagnosis of type 2B von Willebrand's disease (vWD). Analysis involves the use of a synthetic universal heteroduplex generator (UHG). To test the validity of the technique, we have applied UHG screening to seven type 2B vWD patients of previously unknown genotype. Characteristic heteroduplex patterns for Arg543Trp and Val553Met mutations were found in three patients and one patient, respectively. A fifth patient gave a novel pattern and direct sequencing revealed a hitherto unreported candidate mutation (Ser547Phe) 8 bases downstream of an "identifier" deletion in the UHG molecule. The two remaining patients gave normal heteroduplex patterns; an Arg578Gln mutation was identified by PstI digestion in one individual and no mutation could be identified in the sequence covered by the UHG in the final patient. Using a combination of UHG technology and restriction analysis, over 85% of type 2B vWD patients can be rapidly diagnosed by genotype.

Amino Acid Sequence↗

Megakaryocyte growth and development factor and interleukin-3 induce patterns of protein-tyrosine phosphorylation that correlate with dominant differentiation over proliferation of mpl-transfected 32D cells.

Recently, the ligand for c-mpl, a member of the family of cytokine receptors, was cloned and found to be a physiologic regulator of platelet homeostasis. We report that megakaryocyte growth and development factor (MGDF, thrombopoietin [TPO], c-mpl ligand ) induces differentiation in a majority of mpl-transfected 32D cells, while interleukin (IL)-3 is exclusively mitogenic in this system. MGDF differentiation, as measured by decreased proliferation, changes in cellular morphology, increased adherence, and downregulation of very late antigen (VLA)-4, is dominant over IL-3 proliferation. MGDF induces tyrosine-phosphorylation of mpl, JAK2, SHC, SHPTP-1 (HCP, motheaten) and SHPTP-2 (Syp, PTP-1D) within 30 seconds of stimulation, as well as of vav and MAPK with slightly delayed kinetics. A fraction of mpl and JAK2 is preassociated, and the stoichiometry of this complex is unaltered by cytokine stimulation. After MGDF stimulation, we detect interactions among SHC, grb2, SHPTP-1, SHPTP-2, and the mpl/JAK2 complex. IL-3 induces phosphorylation of the above proteins with the exception of mpl and also causes weak JAK1 phosphorylation. Although similar in composition, the MGDF- and IL-3-induced complexes of signal transducers appear to be assembled in different configurations, especially with respect to SHPTP-2. Both MGDF and IL-3 induce tyrosine phosphorylation of STAT3 (APRF) and STAT5 (MGF), with MGDF favoring STAT3 while IL-3 predominantly causes STAT5 phosphorylation. In addition, some proteins become tyrosine-phosphorylated in response to MGDF only, suggesting that we may have detected differentiation-specific signal transducers. These include a number of high-molecular-weight proteins (140 to 200 kD) and one 28-kD protein that becomes tyrosine-phosphorylated only briefly.

Amino Acid Sequence↗

Hepatic granulomas following liver transplantation. Clinicopathologic features in 42 patients.

Liver granulomas have long been known to pose diagnostic problems for pathologists; however, their prevalence and associated etiologic factors have not been studied in liver transplant patients. We reviewed 3632 liver biopsy specimens from 563 patients at two institutions and identified 42 patients with posttransplant granulomas. A possible or probable etiologic factor was identified in 30 (71%) cases. Most were epithelioid granulomas and microgranulomas located in the parenchyma associated with hepatocyte necrosis (21 cases, 50%). Portal-based granulomas were associated with recurrent primary biliary cirrhosis (5 cases, 12%), acute cellular rejection (2 cases, 4.8%), and a foreign body-type reaction (1 case, 2.4%). One case was associated with tuberculosis (2.4%), 4 cases occurred in a fatty liver (9.5%), and 8 patients had liver granulomas but no other significant abnormality. The granulomas were most frequent in the first 7 months after transplantation when the patients were biopsied more often and underwent episodes of rejection or acute hepatitis. Portal-based granulomas in this period were usually associated with acute cellular rejection. After 7 months, the frequency of granulomas as well as the number of biopsies decreased and portal-based granulomas associated with recurrent primary biliary cirrhosis were most common (5 cases, 12%). Rare, late-appearing parenchymal granulomas were also seen (3 cases) and consisted of 1 lipogranuloma and 2 cases of epithelioid granuloma. The latter were thought, in 1 patient, to be associated with parenchymal hepatocyte necrosis; the others were of unknown etiology.

Biopsy↗

Variable expression of CD3-zeta chain in tumor-infiltrating lymphocytes (TIL) derived from renal-cell carcinoma: relationship with TIL phenotype and function.

It has been reported that in mice and in humans, tumor-infiltrating lymphocytes (TIL) may exhibit a defect in CD3-zeta-chain expression. Therefore, the level of CD3-zeta was analyzed in TIL derived from patients with renal-cell carcinoma, and its correlation with TIL phenotype and function was assessed. We identified 4 out of 13 tumor-infiltrating lymphocytes derived from renal-cell carcinoma, with a significant decrease in CD3-zeta-chain expression as compared with control peripheral-blood lymphocytes. This defect was never found after culturing TIL with IL2. In one case, the low expression of zeta chain observed in TIL on day 0 was reversed when TIL were cultured with IL2. The zeta-chain level did not seem to predict the growth of TIL in response to IL2. All the TIL, irrespective of the level of zeta-chain expression, exhibited lower proliferative response when stimulated with PHA or anti-CD3 MAb in comparison with normal peripheral-blood mononuclear cells. Nevertheless, in this limited series of patients, a correlation was observed between the level of zeta-chain expression and T-cell infiltration (p = 0.02). After TIL stimulation with PHA or anti-CD3, in contrast to IL2 or IFN-gamma production, a trend towards a relationship between TNF alpha induction and the level of zeta-chain expression was observed.

CD3 Complex↗

Experimental ablation of outflow tract muscle with a thermal balloon catheter.

BACKGROUND: Pulmonary balloon valvuloplasty has been performed in selected patients with tetralogy of Fallot as an alternative to surgical palliation; this technique is limited, however, by the fact that the balloon has little effect on the dynamic, muscular contribution to outflow tract obstruction. In an experimental model, we used a new thermal balloon catheter to ablate right ventricular outflow tract muscle. We evaluated the acute efficacy and muscle ablation parameters of this technology and its effects after myocardial healing. METHODS AND RESULTS: A prototype electrolyte-filled balloon catheter, heated by radiofrequency energy, was constructed. Studies were conducted to determine the optimum electrolyte solution needed to minimize balloon heating time with an unmodified, commercially available radiofrequency generator. In vivo ablations of right ventricular outflow tract muscle with the thermal balloon were performed in lambs that were divided into three groups (n = 5 each) according to the duration of thermal energy delivery (20, 40, and 60 seconds, respectively). Ablated lesion volume increased (460 +/- 63 to 1156 +/- 256 mm3) as the energy delivery time increased (20 to 60 seconds) and was correlated with delivered energy, temperature integral, and maximum epicardial surface temperature (r = .85, .82, and .72, respectively). All five lesions in the 60-second group showed an acute decrease of the wall thickness. Additional in vivo ablations were performed in 6 animals in which survival studies showed muscle thinning, healing by fibrosis, and no evidence of aneurysm formation. CONCLUSIONS: Thermal energy can be used with a balloon catheter delivery system to ablate myocardium. This study suggests that this energy delivery technology might be useful for relief of muscular outflow tract obstruction and that further studies are warranted.

Animals↗

Norepinephrine infusion following cardiopulmonary bypass: effect of infusion site.

The placement of left atrial catheters following cardiopulmonary bypass (CPB) allows accurate monitoring of left ventricular filling pressures, as well as access for the infusion of vasoactive drugs. While the left atrial administration of norepinephrine (NE) is thought to provide higher systemic arterial NE levels while minimizing any pulmonary vasoconstriction, no study critically compares central venous and left atrial NE infusion following cardiopulmonary bypass. A canine model was used to compare central venous and left atrial NE infusion at three dosages (0.2, 0.4, and 0.6 microgram/kg/min) both prior to CPB and following 2 hr of hypothermic CPB at 27 degrees C. Prior to CPB, there was no difference in the hemodynamic efficacy of central venous and left atrial NE infusion at any dosage. The pulmonary circulation metabolized 16-29% of circulating NE. Only at the 0.2 microgram/kg/min dose was there a difference in the arterial NE level between central venous (3474 +/- 486 pg/ml) and left atrial (5504 +/- 751 pg/ml, P = 0.019) NE administration. Above this dose, no difference in the arterial NE level was identified. Following 2 hr of CPB, the pulmonary endothelium metabolized a significantly higher percentage of circulating NE (35-42%). Despite this increased extent of pulmonary metabolism, there was no difference in the hemodynamic efficacy or the resulting arterial NE level of central venous and left atrial NE infusion at a given dose. In conclusion, the results demonstrate: (1) increased pulmonary NE uptake following CPB, and (2) no difference in hemodynamic efficacy between central venous and left atrial NE infusion over a wide range of three doses in a model with normal lungs.

Animals↗

Systemic activation of 15-lipoxygenase in heart, lung, and vascular tissues by hypercholesterolemia: relationship to lipoprotein oxidation and atherogenesis.

There is evidence that oxidized lipoproteins are a major contributing factor in atherosclerosis. 15-Lipoxygenase is the principal mammalian enzyme that can oxidize polysaturated fatty acids present in intact lipoproteins, and in membrane phospholipids in situ. We, and others, have reported previously that levels of the enzyme are increased in aortas of cholesterol-fed and spontaneously atherosclerotic WHHL rabbits. In the present study, rabbits were fed an atherogenic diet containing 1% cholesterol for 14 weeks, and levels of [14C]arachidonate metabolizing enzymes in the excised tissues were measured by HPLC analysis. 15-Lipoxygenase levels in heart, aortic adventitia, and lung, but not in liver, were increased up to 100-fold above controls, without major significant changes in prostaglandin endoperoxide synthases or the 5- and 12-lipoxygenases. The induced 15-lipoxygenase activity in the aortic adventitia was approximately 15 times greater than that found in the vessel wall. Hypercholesterolemia and elevated 15-lipoxygenase were associated with a 40% lowering of blood hematocrit. The hemolytic agent phenylhydrazine duplicated the effects of hypercholesterolemia on hematocrit, and induced up to 100-fold increases in 15-lipoxygenase activity in tissues within 7 days. The induced 15-lipoxygenase activities in heart and lung were 4 and 8 times greater, respectively, than in reticulocytes, previously the richest known source of the enzyme. Direct measurements of hemoglobin content also demonstrated that contaminating reticulocytes were not the source of the tissue enzyme. A similar tissue-specific activation of 15-lipoxygenase was observed in rat heart and lung, but also not in liver. It is concluded that the elevated level of 15-lipoxygenase activity previously reported in atherosclerotic aorta is symptomatic of a generalized and massive induction of the enzyme in cardio-pulmonary tissues by hypercholesterolemia, which may be related to the membrane perturbation and increased hemolysis that is induced by cholesterol feeding.

Animals↗

Limitations of the efficacy of hemoglobin-based oxygen-carrying solutions.

Improvements in hemoglobin-based oxygen-carrying (HBOCs) solutions have overcome the toxicities that plagued earlier efforts. However, limitations of the efficacy of HBOCs are emerging. The potential limitations of an HBOC were studied in an ovine model (n = 6) of exchange transfusion. Hemodynamic, oxygen transport, and hemoglobin kinetic parameters were examined during isovolumic blood exchange to a final hematocrit of 3.2 +/- 0.7% and a plasma hemoglobin concentration of 8.1 +/- 0.4 g/dl while sheep were awake and breathing room air. However, the infusion of HBOC was associated with immediate increases in systemic and pulmonary arterial pressures. Despite hemodilution with HBOC, systemic and pulmonary vascular resistance increased 43.9% (P < 0.001) and 204.2% (P < 0.001), respectively, after HBOC infusion. After blood exchange, the plasma hemoglobin level exhibited a circulatory half-life of 52.7 +/- 18.0 h. The formation of methemoglobin was significant, accounting for 33.0 +/- 7.1% of the total circulating plasma hemoglobin at 24 h; the half-life of HBOC capable of carrying oxygen was 30.1 +/- 5.4 h. This relatively short period of oxygen-carrying efficacy and the observed vasoconstriction properties may constrain the potential applications of HBOC solutions.

Animals↗

Star volumes of villi and intervillous pores in placentae from low and high altitude pregnancies.

Histological sections of placentae from pregnancies completed at low altitude (400 m) and high altitude (3600 m) in Bolivia were analysed using a stereological estimator of the star volumes of villous 'domains' and intervillous 'pores'. The purpose was to test whether or not differences in the overall volumes of these compartments are accompanied by changes in their geometrical relationships. Whilst total placental volume did not vary with altitude, the total volume of villi declined by about 25% and total intervillous volume increased by 40% at high altitude. The star volume of villi also decreased by 25% (from 1.5 x 10(6) microns 3 at low altitude to 1.1 x 10(6) microns 3 at high altitude) whilst the star volume of intervillous pores increased 4-fold (from 87 x 10(6) microns 3 to 461 x 10(6) microns 3). These figures imply that villous domains decrease in size but may be constant in number. The most likely explanation is that villous trees at high altitude are scaled-down versions of their low-altitude counterparts. By contrast, although the intervillous pores enlarge they may decrease in number in the highland organ. This may reflect a change in the number of maternal cotyledons or in the spatial arrangement of villous trees.

Altitude↗

Assessing the safety of pediatric laparoscopic surgery.

We studied deviations from normal physiology in piglets (n = 10; average weight 5.75 kg) during carbon dioxide (CO2) pneumoperitoneum. Cardiopulmonary data were gathered during varying intraabdominal pressures (IAP = 8, 12, 15, 20 mm Hg), each sustained for 10 mins. Each animal was its own preinsufflation and exsufflation control. A rapid, significant rise in arterial CO2 pressure from preinsufflation (46.5 +/- 6.7 mm Hg) to insufflation at 20 mm Hg (72.9 +/- 15 mm Hg; p < 0.05) initiated further cardiac adjustments. Responses included a sustained increase in cardiac index (presufflation = 3.1 +/- 1.4; 20 mm Hg IAP = 3.6 +/- 1.2), increased heart rate (preinsufflation = 121 +/- 21; 20 mm Hg IAP = 150 +/- 28; p < 0.05), and left ventricular stroke work (20 mm IAP = 22.7 +/- 8.9; exsufflation 20 min = 15.3 +/- 9.4 g.m/m2; p < 0.05). There was a significant arterial-end CO2 tidal difference throughout insufflation, as great as 15 mm Hg (p < 0.05), suggesting increasing ventilation dead space. Core temperature decreased significantly from preinsufflation (35.3 +/- 1.3 degrees C) to 20 mm Hg IAP (33.6 +/- 1.5 degrees C, p < 0.05). We suggest the following guidelines based on the above data: (a) preoperative examination screening for cardiopulmonary abnormalities; (b) fluid replacement to normal hydration only; (c) cuffed endotracheal tubes for effective ventilation; (d) careful adjustment of minute ventilation to achieve normocapnia; (e) CO2 warming; (f) maximal insufflation pressure of 12 mm Hg; (g) postoperative care emphasizing respiratory and thermoregulation status.

Abdomen↗

Human fertility and population equilibrium.

In preindustrial settings, biological, economic, and social mechanisms caused fertility to respond to feedback signals from population size variation in such a way as to steer populations toward equilibrium levels. It is unlikely that the relevant signal was population density; more plausibly the signal was economic well being, which in turn responded to population density, among other influences. Numerous empirical studies provide persuasive evidence for such a fertility response. However, the response was relatively weak, so that populations responded only slowly to shocks. The half life of a shock-induced deviation from equilibrium was probably on the order of 70 years. Dynamic behavior of population size was discernibly affected only in the long run. Because of the presence of environmental external costs to childbearing, and because fertility response mechanisms were not necessarily intentional, the equilibrium population size and corresponding welfare need not have possessed any desirable properties. In developed countries, these equilibrating mechanisms are no longer in effect.

Animals↗