Automated method for L-carnitine determination

Because of renewed interest in a possible connection between carnitine, lipid disorders, and myopathy, an automated method of analysis is desirable. Deproteinization of serum by use of membrane filter cones and automated assay with a bichromatic analyzer (the ABA-100) substantially increases efficiency without sacrificing the specificity and accuracy of the original manual enzymatic method. The described procedure allows for analysis of 80 speciments a day and is thus suitable for screening of selected populations. Normal values found in blood sera of adults were in the range of 25.0-73.8 mu mol/liter and the method has sufficient sensitivity to accurately measure concentrations as small as 10 mu mol/liter.     

Urography during pregnancy

A review of 22,971 pregnancies from 1969-74 reveals that the reasons for requesting excretory urography were renal colic of pregnancy 11, pyelonephritis 8, renal contusion 2, threatened abortion 1, and suspected degeneration of fibroid 1. The final diagnoses were similar except that two patients with torsion of ovarian cysts and a renal calculus were discovered. The only intravenous pyelographic examination that was decisive for diagnosis was in a patient with a ureteric calculus. Careful clinical correlation should reduce excretory urography during pregnancy.     

A general method for isolation of high molecular weight DNA from eukaryotes

A new method for isolation of high molecular weight DNA from eukaryotes is presented. This procedure allows preparation of DNA from a variety of tissues such as calf thymus or human placenta and from cells which were more difficult to lyse until now (e.g. Crypthecodinium cuhnii, a dinoflagellate). The DNA obtained in such a way has an average molecular weight of about 200 X 10(6) d and contains very few, if any, single strand breaks.     

Recurrent urothelial tumors following surgery for transitional cell carcinoma of the upper urinary tract

A retrospective analysis of 74 cases of transitional cell carcinoma of the renal pelvis and ureter treated at this institution over the past 30 years is presented. When nephrectomy alone or incomplete nephroureterectomy was performed, subsequent transitional cell carcinoma developed in 30% of the ureteral stumps. Subsequent bladder carcinoma occurred in 25% of the patients with primary upper urinary tract carcinoma. The type of initial surgery performed did not appear to influence this incidence of subsequent bladder tumors. Contralateral upper urinary tract carcinoma developed in only one patient. When nephroureterectomy is performed for carcinoma of the renal pelvis and ureter, a cuff of bladder that includes the ureteral orifice should be removed to obviate recurrent disease in the ureteral stump. Since single-incision nephroureterectomy did not include the intramural ureter in 50% of the cases in which it was performed, a second incision may be required for adequate exposure.     

Operative correction of pectus excavatum: an evolving perspective

Volatile free fatty acids (VFA) in blood increased approximately twofold in dogs subjected to total hepatectomy. The average total plasma VFA preoperatively was 1,585 mug percent and shortly before death, postoperatively, was 2,798 mug percent. The corresponding red cell concentrations were essentially the same. Acetic acid was 81 percent of the total VFA, propionic acid 7 percent, isobutyric acid 4 percent, butyric acid one percent, and isovaleric acid 5 percent. There was little or no isovalerate in red cells. The increments in the individual fatty acids after hepatectomy were highly variable, but the average increase with time was almost linear. The increase in VFA probably reflects an increased utilization of the branched-chain aminoacids by extrahepatic tissues.     

Protein thermostability above 100 degreesC: a key role for ionic interactions

The discovery of hyperthermophilic microorganisms and the analysis of hyperthermostable enzymes has established the fact that multisubunit enzymes can survive for prolonged periods at temperatures above 100 degreesC. We have carried out homology-based modeling and direct structure comparison on the hexameric glutamate dehydrogenases from the hyperthermophiles Pyrococcus furiosus and Thermococcus litoralis whose optimal growth temperatures are 100 degreesC and 88 degreesC, respectively, to determine key stabilizing features. These enzymes, which are 87% homologous, differ 16-fold in thermal stability at 104 degreesC. We observed that an intersubunit ion-pair network was substantially reduced in the less stable enzyme from T. litoralis, and two residues were then altered to restore these interactions. The single mutations both had adverse effects on the thermostability of the protein. However, with both mutations in place, we observed a fourfold improvement of stability at 104 degreesC over the wild-type enzyme. The catalytic properties of the enzymes were unaffected by the mutations. These results suggest that extensive ion-pair networks may provide a general strategy for manipulating enzyme thermostability of multisubunit enzymes. However, this study emphasizes the importance of the exact local environment of a residue in determining its effects on stability.     

Quantitation of vancomycin and its crystalline degradation product (CDP-1) in human serum by high performance liquid chromatography

Mantle cell lymphoma (MCL) patients represent a difficult problem, sometimes to establish the diagnosis but mostly because of their refractoriness to standard lymphoma treatments. Which treatments to apply and to whom is not yet defined. In this study, we attempted to analyze the clinical features, to identify the major prognostic factors, and to evaluate the outcome of 121 MCL patients treated in our institution between 1979 and 1997. Clinical data, treatment modalities, and International Prognostic Index (IPI) score were evaluated. Median age was 63 years. Patients usually presented with advanced stage disease (87%), disseminated lymph nodes (57%), bone marrow involvement (79%), but with a good performance status (PS) (81%). Lymphocytosis >4000/microl and/or peripheral blood involvement was present in 36% of cases, and gastrointestinal disease in 18%. The t(11;14)(q13;q32) and/or bcl-1 rearrangement was detected in 47/57 studied cases. Median overall survival (OS) was 3.12 years and a longer survival was significantly associated with younger age (<70 years), good PS (<2), localized disease (stage I-II), fewer than two extra-nodal sites, absence of spleen or peripheral blood involvement, normal serum LDH and beta2-microglobulin levels, and hemoglobin level greater than 12 g/dl. However, the IPI failed to identify patients with longer OS and in a multiparametric analysis, only older age, hemoglobin less than 12 g/dl, poor PS, and blood involvement were associated with a poorer outcome. Treatment modalities had no impact on survival with 75% of patients relapsing or progressing. Our data showed that the poor outcome of MCL patients is mainly related to adverse patient characteristics, a highly disseminated tumor, and some unknown parameters associated with the refractoriness to standard therapy.     

NO/cGMP pathway is involved in exocrine secretion from rat pancreatic acinar cells

OBJECTIVE: To identify factors associated with the initiation of breast-feeding in a predominantly Puerto Rican population living in inner-city Hartford, Conn. DESIGN: Retrospective study of 144 Latino women (mean +/- standard deviation age = 26.3 +/- 5.7 years) with children at least 1 year old but younger than 6 years old (mean +/- standard deviation age = 3.0 +/- 1.2 years) at the time of the survey. Women were recruited from agencies sponsoring health programs for mothers and children. They were interviewed in their homes (69%) or at the Hispanic Health Council, Hartford, Conn (31%). SUBJECTS/SETTING: Low-income Latino women who had at least 1 preschooler at the time of the interview. The women lived in inner-city Hartford, and the overwhelming majority were Puerto Rican and received welfare assistance and food stamps. Seventy-eight percent of the women chose to be interviewed in Spanish; the other 22% were interviewed in English. STATISTICAL ANALYSES: Explanatory variables that related to breast-feeding initiation (P < or = .2) in bivariate chi 2 analyses were entered into a multivariate logistic regression model that was reduced using backward stepwise elimination procedures. RESULTS: Multivariate analyses indicated that breast-feeding the previous child, shorter length of maternal residence in the United States, not receiving prenatal bottle-feeding advice, more recent birth, and higher birth weight were positively associated with breast-feeding initiation. A major reason for choosing not to breast-feed was that women felt socially uncomfortable doing it. APPLICATIONS: Breast-feeding initiation was more likely in Latino women who received prenatal breast-feeding counselling and postpartum support. Mothers of low-birth-weight infants and women breast-feeding for the first time may need additional help. These findings can be used by programs like the Special Supplemental Nutrition Program for Women, Infants, and Children to increase breast-feeding initiation.     

Optimization of flow in additively manufactured porous columns with graded permeability

Chemical engineering systems often involve a functional porous medium, such as in catalyzed reactive flows, fluid purifiers, and chromatographic separations. Ideally, the flow rates throughout the porous medium are uniform, and all portions of the medium contribute efficiently to its function. The permeability is a property of a porous medium that depends on pore geometry and relates flow rate to pressure drop. Additive manufacturing techniques raise the possibilities that permeability can be arbitrarily specified in three dimensions, and that a broader range of permeabilities can be achieved than by traditional manufacturing methods. Using numerical optimization methods, we show that designs with spatially varying permeability can achieve greater flow uniformity than designs with uniform permeability. We consider geometries involving hemispherical regions that distribute flow, as in many glass chromatography columns. By several measures, significant improvements in flow uniformity can be obtained by modifying permeability only near the inlet and outlet.