Resolving isomeric peptide mixtures: a combined HPLC/ion mobility-TOFMS analysis of a 4000-component combinatorial library.

A reversed-phase high-performance liquid chromatography (HPLC) separation approach has been combined with ion mobility/time-of-flight (TOF) mass spectrometry in order to characterize a combinatorial peptide library designed to contain 4000 peptides of the general form NH2-Xxx-Xxx-XXX-CO2H, NH2-Ala-Xxx-Xxx-Xxx-CO2H, NH2-Ser-Ala-Xxx-Xxx-Xxx-CO2H and NH2-Leu-Ser-Ala-Xxx-Xxx-Xxx-CO2H (where Xxx represents a randomization over 10 different amino acids: Ala, Arg, Asp, Glu, Gly, Leu, Lys, Phe, Ser, and Val). Addition of the gas-phase mobility separation between the HPLC separation and TOF measurement dimensions makes it possible to resolve many peptide isomers that have identical retention times (and masses).     

ESI/ion trap/ion mobility/time-of-flight mass spectrometry for rapid and sensitive analysis of biomolecular mixtures

An ion trap/ion mobility/time-of-flight mass spectrometry technique is shown to be a rapid and sensitive means of analyzing peptide/protein mixtures. In this approach, an ion trap is used to accumulate ions that have been electrosprayed from a mixture into concentrated packets. The ion packets are injected into a drift tube where components of the mixture are separated based on differences in mobility through a buffer gas. Ions that exit the drift tube are dispersed in a time-of-flight mass spectrometer for mass-to-charge (m/z) determination. The gas-phase separation strategy reduces congestion in the mass spectrum, and experimental mobilities complement m/z measurements in assigning peaks. Examples of the application of the approach to identification of peptides (from tryptic digests) and to separation of charge-state distributions from electrospray of a mixture containing ubiquitin and myoglobin are presented. Most peptides that are observed from tryptic digests of proteins such as cytochrome c and myoglobin can be identified from data that are acquired in under 1 min; studies of mixtures with known compositions indicate that detection limits are approximately 0.5-3 pmol for individual components. Factors that may influence the distributions that are observed, such as storage time in the trap, injection voltages used for the mobility experiment, and variations in ion cross section with charge state, are discussed.     

Academic associate program: integrating clinical emergency medicine research with undergraduate education

MATERIALS AND METHODS: The diagnostic accuracy of scintimammography with 99mTc-MDP was evaluated in 400 consecutive women with clinical or mammographic suspicion of breast cancer, candidate to surgery and/or excisional biopsy. Lateral views of both glands were acquired, in prone position, 5-10 min after the injection of 550-740 MBq of 99mTc-MDP. The scintigraphic results were compared to mammograms and classified using the histological findings as gold standard. RESULTS: Mammography was suggestive for cancer in 231 (57%), suspicious in 49 (12%) and indeterminate in 120 (31%) patients. Breast carcinoma was histologically proven in 330 women, benign breast diseases in 70. The tumor size ranged from 4 x 5 to 50 x 60 mm. 99mTc-MDP visualized as foci of increased uptake 305/330 cancers (92%). In particular, in women with indeterminate mammograms the SMM had a diagnostic accuracy of 84%, correctly characterizing 101/120 lesions. Twenty missed cancers had largest diameter < or = 10 mm, 5 < or = 15 mm. Lack of 99mTc-MDP uptake occurred in 64 out of 70 benign lesions. These lesions were classified as truly negative. Conversely, 3 fibroadenoma and 3 epithelial hyperplasia with moderate or severe atypia were falsely positive. The overall specificity was 91.5%; the accuracy was 92%, the positive and negative predictive values were respectively 98% and 72%. CONCLUSIONS: The results obtained in this study suggest that 99mTc-MDP scintimammography accurately detects breast carcinomas with largest diameter > 10 mm; it differentiates malignant from benign lesions, and it shows promising insights in characterizing breast abnormalities mammographically indeterminate.     

Disposition of butadiene epoxides in Sprague-Dawley rats

1,2-Epoxybutene (BMO) and diepoxybutane (BDE) are metabolic products of 1,3-butadiene in rodents. Both BMO and BDE are suspect in the development of tumors in rats and mice. To understand the distribution and elimination of these compounds in the absence of the rate-limiting production from butadiene, the pharmacokinetics of BMO and BDE in blood were determined in adult male Sprague-Dawley rats following intravenous administration. All animals were dually cannulated in these studies. For the BMO studies, rats were dosed with 71, 143, or 286 mumol/kg BMO (n = 3 for each dose group). For the BDE studies, rats were dosed with 523 mumol/kg BDE (n = 3). All animals tolerated the BMO and BDE doses without grossly observable adverse effects. Blood was drawn at predetermined time points and extracted in methylene chloride. BDE and BMO concentrations were quantitated by gas chromatography or gas chromatography/mass spectrometry. The BMO distribution half-lives were short and ranged from 1.4 min at the lowest dose to 1.8 min at the highest dose. Volume of distribution at steady state ranged from 0.53 +/- 0.17 to 0.59 +/- 0.31 l/kg. Systemic clearances ranged from 67 +/- 17 to 114 +/- 20 ml/min per kg. The terminal elimination half-lives were also short and ranged from 5.7 to 8.5 min among the doses. The pharmacokinetic parameters after an i.v. dose of 523 mumol/kg BDE were a distribution half-life of 2.7 min, terminal elimination T1/2 of 14 min, volume of distribution at steady state of 0.73 +/- 0.06 l/kg, and systemic clearance of 76 +/- 8 ml/min per kg. These pharmacokinetic parameters demonstrate the similarity between disposition of the two epoxides in rats, that include a rapid distribution after i.v. administration into a small extravascular body compartment as well as a rapid elimination from blood. These pharmacokinetic data provide useful blood clearance information for assessing the critical physiological and biochemical determinants underlying the disposition of butadiene epoxides.     

Effect of influenza immunization on immunologic and virologic characteristics of pediatric patients infected with human immunodeficiency virus

In this paper, a data-parallel computer is used to provide the memory and reduction in computer time for solving large finite-difference bidomain problems. The finite-difference grid is mapped effectively to the processors of the parallel computer, simply by mapping one node to one (virtual) processor. Implemented on the connection machines (CM's) CM-200 and CM-5, the data-parallel finite-difference algorithm has allowed the solution of finite-difference bidomain problems with over 2 million nodes. Details on the algorithm are presented together with computational performance results.     

The use of different fixatives and hydrophilic embedding media (Historesin and Unicryl) for the study of embryonic tissues

As part of a research project exploring ways through which we can understand the crying infant and its family, this study focuses on the experiences of fathers during labour and delivery of their infant. In a previous part of the project it was shown that fathers' negative experiences during the childbirth were correlated with the amount of crying in the infant during the first months after birth. The aim of the present study was to explore and interpret the experiences that fathers reported in an interview when the infant was between six months and one year of age. A hundred and nine fathers were interviewed. The interviews, which took place in the families' homes and with both parents present, were carried out in dialogue form with open-ended questions. The results reveal that complications during the delivery were significantly correlated with the amount of crying in the infant. Feelings of helplessness, of guilt and that staff behaviour had been negative were more common in the group of fathers who experienced the delivery as a negative event. 'Locus of control' seems to be the most relevant concept.     

Prevalence of thinness and excessive fatness in a group of school children of the city of Cordoba, Argentina

Findings of a nutritional evaluation study, using triceps skinfold, are reported. The study was carried out in 1,615 school-children from 5 to 12 years, pertaining to the low socioeconomic status of the city of Córdoba, Argentina. Measurements were done during the years 1983 and 1984 by Center's examiners with a Lange caliper, according to international anthropometric methodology. Interintra observer technical error was found to be within tolerance limits reported by other authors. Local standards were used to determine the prevalence of thinness and excessive fatness, by comparing them to tricep skinfold of each child, and selecting those children where this parameter presented values less than or equal to 10th percentile and fatness greater than or equal to 90th percentile. Furthermore, comparison between median triceps skinfold of the examined group, the local standards and Frisancho's norms for US population was also made. Results revealed that: a) the prevalence of thinness (19.9%) was almost twice the percentage expected for a normal population; in contrast, occurrence of excessive fatness (6.4%) was found to be below the expected value; b) prevalence of thinness was not statistically associated to age nor sex; c) frequency of excessive fatness was significantly higher in the 8-11 year-old male group (8, 9%, p much less than 0.0005) and in the girls group (10.8%), P much less than 0.005) the prevalence of fatness in girls increased with age, and figures revealed that from eight years onwards this exceeded the expected percentage, and d) median triceps skinfold of the group under study was generally below local and foreign norms. It is concluded that potential or real nutrition problems as those mentioned above, may be grossly identified using the triceps skinfold as the single anthropometric indicator.     

The human erythroid-specific transcription factor EKLF localizes to chromosome 19p13.12-p13.13

Erythroid Krüppel-like factor (EKLF) is a trans-acting factor that binds specifically to the beta-globin promoter CACCC box. EKLF is essential for adult erythroid development in mice. We have cloned and sequenced a cDNA encoding the human homologue of this gene, which shares 69% identity with the mEKLF protein. The expression of hEKLF is restricted to fetal liver and adult bone marrow. The genomic locus of hEKLF was mapped to chromosomal band 19p13.12-p13.13, using fluorescence in situ hybridization.     

Failure of colonoscopic surveillance in ulcerative colitis

A prospective surveillance programme for patients with longstanding (> = 8 years), extensive (> = splenic flexure) ulcerative colitis was undertaken between 1978 and 1990. It comprised annual colonoscopy with pancolonic biopsy. One hundred and sixty patients were entered into the programme and had 739 colonoscopies (4.6 colonoscopies per patient; 709 patient years follow up). Eight eight per cent of examinations reached the right colon. There was no procedure related death. One Dukes's A cancer was detected. Forty one patients (25%) defaulted. Of these 25 remain well; 13 are unaccounted for, and one died from colonic cancer. One patient had colectomy for medical reasons, and another died of carcinoma of the pancreas. Retrospectively an additional 16 eligible patients were identified who had not been recruited. Of these, 14 remain well, two are unaccounted for. None developed colonic cancer. Four patients refused colonoscopy. All remain well. Over the same period seven other cases of colonic cancer were found in association with ulcerative colitis, two in patients who had erroneously been diagnosed as having only proctitis and were therefore not entered into the programme, but were found at operation to have total colitis, one in a patient with colitis of seven years duration, and four patients who had previously attended the clinic but had been lost to follow up before 1978 and then had represented with new symptoms during the surveillance period. Thus, of the nine colitis related cancers diagnosed in this centre during the study period only one was detected by the surveillance programme. The results of this large study, a a review of published works, cast doubts on the effectiveness of colonoscopic surveillance programmes in detecting colorectal cancer in patients with ulcerative colitis.     

Large Grinding Mill Drives Update

The trend to larger grinding mills with the attendant increase in motor drive horsepower has affected the options for drive equipment. Also the development of large adjustable speed drives has created new possibilities for mill drives. Grinding mill torque characteristics, drive requirements, and torsional excitations of the various types of available drives will be briefly reviewed.