Repair of DNA methylphosphotriesters through a metalloactivated cysteine nucleophile

The Escherichia coli Ada protein repairs methylphosphotriesters in DNA by direct, irreversible methyl transfer to one of its own cysteines. Upon methyl transfer, Ada acquires the ability to bind specific DNA sequences and thereby to induce genes that confer resistance to methylating agents. The amino-terminal domain of Ada, which comprises the methylphosphotriester repair and sequence-specific DNA binding elements, contains a tightly bound zinc ion. Analysis of the zinc binding site by cadmium-113 nuclear magnetic resonance and site-directed mutagenesis revealed that zinc participates in the autocatalytic activation of the active site cysteine and may also function as a conformational switch.     

Detection rates for abnormal cervical smears: what are we screening for?

We analysed rates of detection for smear abnormalities in 255,000 women served by the Bristol screening programme. The programme began in 1966 with the aim of eradicating the 30-40 deaths each year in Bristol from cervical cancer. Organisation has been good and population uptake has been high for the past 15 years. Records were computerised in 1977. During the 1988 to 1993 screening round, 225,974 women were tested. New smear abnormalities were found in 15,551, of whom nearly 6000 were referred for colposcopy. These numbers are excessively high in comparison with the incidence of the malignancy we are trying to prevent. The effect of screening on death rates in Bristol is too small to detect. Our conclusion is that despite good organisation of the service, much of our effort in Bristol is devoted to limiting the harm done to healthy women and to protecting our staff from litigation as cases of serious disease continue to occur. The real lesson from 30 years' cervical screening is that no matter how obvious the predicted benefit may seem for any screening test, introduction should never take place without adequate prior evaluation of both positive and negative effects in controlled trials.     

Participation in continuing professional education in nursing: findings of an interview study

The evidence suggests that enrolled nurses and those working part-time and on night duty consistently attend less continuing professional education than their more senior, full-time and day duty colleagues. This was substantiated in the findings of this study which also highlighted the paucity of opportunities for continuing education among unqualified practitioners. In the light of recent developments within the health services, this inequitable provision of continuing professional education to all nursing practitioners will need to be addressed and resolved. Continuing professional education provided on an arbitrary and random basis will not afford maximum positive outcomes for clients, the service and practitioners and has the potential to increase the levels of frustration among disadvantaged groups of nursing staff.     

Enzymatic preparation of heparin oligosaccharides containing antithrombin III binding sites

Two new oligosaccharides were prepared from heparin by its partial depolymerization using heparin lyase I (EC 4.2.2.7) in an attempt to prepare oligosaccharides having intact antithrombin III binding sites. The oligosaccharides were purified by chromatography on the basis of both size and charge and demonstrated a high level of purity by capillary electrophoresis. One- and two-dimensional 1H NMR spectroscopy at 500 MHz revealed the structure of each oligosaccharide. The octasaccharide and decasaccharide are DeltaUAp2S(1-->4)-alpha-DGlcNpS6S(1-->4)-alpha-L-IdoAp (1-->4)-alpha-D -GlcNpAc6S(1-->4)-betaD-GlcAp(1-->4)-alpha-D-GlcNpS 3S6S(1-->4)-alpha- L-IdoAp2S(1-->4)alpha-D-GlcNpS6S (where DeltaUAp is 4-deoxy-alpha-L-threo-hex-enopyranosyluronic acid, GlcNp is 2-amino-2-deoxy-glucopyranose, GlcAp is glucopyranosyluronic acid, S is sulfate and Ac is acetate) and DeltaUAp2S(1-->4)-alpha-D-GlcNpS6S(1-->4)-alpha-L-IdoAp++ +(1-->4)-alpha- D-GlcNpAc6S (1-->4)-beta-D-GlcAp(1-->4)-alpha-D-GlcNpS3S6S(1-->4)-alpha- L-IdoAp2S (1-->4)-alpha-D-GlcNpS6S(1-->4)-alpha-L-IdoAp2S(1-->4)-alpha -D-GlcNpS 6S, respectively. A hexasaccharide containing a similar structural motif to that found in the antithrombin III binding site and having greatly reduced anticoagulant activity was also isolated. The structure of the hexasaccharide is DeltaUAp2S(1-->4)-alpha-D-GlcNpAc6S(1-->4)-beta-D-GlcAp++ +(1-->4)-alpha- D-GlcNpS3S6S(1-->4)-alpha-L-IdoAp(1-->4)-alpha-D-GlcNpS6S . The octasaccharide and decasaccharide correspond to the predominant structural motif found in porcine intestinal mucosal heparin. Sufficient quantities of the decasaccharide were obtained to examine its interaction with antithrombin III using microtitration calorimetry. This decasaccharide bound to antithrombin III with similar avidity as heparin and showed comparable anticoagulant activity, as determined using an antithrombin III dependent anti-factor Xa assay. Interestingly, while both decasaccharide and heparin bound to antithrombin with nanomolar affinity, very little heat of binding was observed.     

The heterogeneity of cell populations by the cytochemical characteristics of the cellular chromatin

Cell heterogeneity of cytochemical characteristics of chromatin in the norm and after provocative factors in vitro (a heating at 45 degrees C, X-irradiation and joint actions of heating and X-ray irradiation) was investigated in the peripheral blood mononuclear cells of sheep and mouse, the lymphoid cell populations of the murine bone marrow, spleen and thymus. As a criterion of cytochemical heterogeneity were used the distinctions in the rate of staining of cell nuclei with DNA-specific dyes, that was registered by means of measurement of coefficient variation of the DNA-histogram basis peak, with a flow cytometry. Heterogeneity of cytochemical characteristics are dealt with showed a variability of spatial organization of interphase nuclei of single cells. The provocative factors bring about a decrease in the level of the cytochemical characteristics heterogeneity in the sheep mononuclear cells. In the murine cell populations studied no such equalizing of the marker of variability was revealed.     

Stability of systems with several monotone nonlinearities

This paper is devoted to stability problems of automatic control systems. Frequency domain criteria for global asymptotic stability of equilibrium sets for new classes of nonlinearities are derived. Results are illustrated by examples.     

Ultrasonographically-guided fine-needle aspiration cytology in the diagnosis of colonic lesions

BACKGROUND: The use of fine-needle aspiration cytology (FNAC) in the diagnosis of colonic lesions was investigated. METHODS: Some 22 patients (median age 71 years) with a colonic lesion identified on abdominal ultrasonography underwent ultrasonographically-guided FNAC using a 21-G needle. The sample was checked immediately by a cytopathologist for adequacy. RESULTS: Eighteen patients had colonic carcinoma; aspiration cytology detected malignant epithelial cells consistent with colonic carcinoma in 17 patients and severely dysplastic cells in one patient. The sensitivity and specificity of ultrasonographically-guided FNAC in the diagnosis of colonic carcinoma was 94 and 100 per cent respectively. The remaining four patients had a diagnosis of ileocaecal tuberculosis, ileocaecal Crohn's disease, and metastatic adenocarcinoma in the liver with no identifiable primary (two patients). One demonstrated granulomata, grew acid-fast bacilli and the patient was treated for tuberculosis. One had inflammatory cells and the patient was found to have Crohn's disease on histology. The remaining two patients had confirmed metastatic adenocarcinoma in the liver on aspiration cytology but suspected colonic lesions were found to be benign on cytological examination and no primary lesion was subsequently demonstrated. There were no complications of FNAC and patients complained of minimal discomfort. There has been no evidence of tumour recurrence with a median follow-up of 12 (range 1-25) months. CONCLUSION: Ultrasonographically-guided FNAC is a valid method for the diagnosis of colonic tumours.