Crystal structure of CcdB, a topoisomerase poison from E. coli

The crystal structure of CcdB, a protein that poisons Escherichia coli gyrase, was determined in three crystal forms. The protein consists of a five-stranded antiparallel beta-pleated sheet followed by a C-terminal alpha-helix. In one of the loops of the sheet, a second small three-stranded antiparallel beta-sheet is inserted that sticks out of the molecule as a wing. This wing contains the LysC proteolytic cleavage site that is protected by CcdA and, therefore, forms a likely CcdA recognition site. A dimer is formed by sheet extension and by extensive hydrophobic contacts involving three of the five methionine residues and the C terminus of the alpha-helix. The surface of the dimer on the side of the alpha-helix is overall negatively charged, while the opposite side as well as the wing sheet is dominated by positive charges. We propose that the CcdB dimer binds into the central hole of the 59 kDa N-terminal fragment of GyrA, after disruption of the head dimer interface of GyrA.     

Dietary practices of South African ultradistance runners

Training (T) and prerace (PR) dietary intakes of male and female athletes participating in a 90-km ultramarathon and the usual diets of matched, sedentary controls were investigated using 24-hr dietary records. Supplement use, mean weekly training distance, and race performance times were recorded. Macro- and micronutrient intakes were analyzed using computerized nutritional analysis programs. Total mean energy intake in the T and PR diets of the runners was 10.1 and 12.8 MJ in the men (n = 150) and 7.5 and 9.1 MJ in the women (n = 23). Mean relative contribution of CHO to the runners' total kilojoule intake increased from 50.0 and 49.5% in the T diets to 57.7 (p < .05; n = 153) and 56.4% (p < .05; n = 23) in the PR diets of male and female runners, respectively, and energy-boosting supplements were included in the PR diets of 48% of female and 59% of male runners. Seventy-eight percent of female and 62% of male runners used vitamin and mineral supplements in their T diets as opposed to 39% of female and 28% of male controls. No statistically significant relationship was found between total kilojoule, CHO, fat, protein, and selected micronutrient intake during the 3 days before the race and performance in the 90-km event in runners of homogenous training status and gender.     

High frequency of neoplasia in patients with autoantibodies to centromere protein CENP-F

OBJECTIVE: To study the clinical features of patients with autoantibodies to centromere protein CENP-F and the frequency of CENP-F autoantibodies in patients with various diseases. DESIGN: Retrospective clinical and serologic study. METHODS: Thirty-six patients with anti-CENP-F were identified by a characteristic pattern of indirect immunofluorescence (IIF) on HEp-2 cells. Fifty patients with melanoma, 50 with breast cancer, 10 with lung cancer, 354 with systemic sclerosis, 120 with systemic lupus erythematosus and 50 with rheumatoid arthritis were also studied. Recombinant proteins were produced from 5 CENP-F cDNA clones representing amino acids 2192-3317 (p-F1), 5561-7126 (p-F2), 5892-6883 (p-F3), 7538-10,116 (p-F4) and 9242-10,096 (p-F5). The presence of CENP-F antigen was studied in a breast carcinoma cell line, cryosections of breast carcinoma, normal breast tissue and tonsils. RESULTS: Twenty-two of 36 patients with CENP-F antibodies had neoplasms; breast (9/22) and lung (5/22) cancer were the most common diagnoses. Thirty-three sera were available for further study; when tested for reactivity to the recombinant peptides, the sera of 21 of 21 patients with neoplasms and 5 of 12 patients with other diseases bound the C-terminal p-F4 peptide. When the terminal third of the p-F4 peptide (p-F5) was studied, a significant difference in pattern of reactivity was not detected. By comparison, the frequency of reactivity with peptides representing other domains of CENP-F was less than that with p-F4 (p-F2 > p-F3 > p-F1). CENP-F autoantibodies were not found in any of the control sera from patients with systemic lupus erythematosus, rheumatoid arthritis or systemic sclerosis or in unselected sera from various malignancies. CENP-F antigens were identified in breast carcinoma tissue but were rarely observed in normal tissues. CONCLUSIONS: A high proportion of individuals with CENP-F antibodies have neoplasia, and there is a bias among their sera for reactivity with determinants in the carboxy terminal domain of CENP-F. CENP-F antigens appear to be highly expressed in malignant tissues.     

Dimensions of care in five United States nursing homes: identifying invisible work in care-giving

OBJECTIVE: We recently reported that acupuncture treatment of stroke patients in the subacute stage gave additive therapeutic benefit. The purpose of the present study was to determine, approximately one year after discharge from the rehabilitation hospital, whether the group differences still remained. DESIGN: The patients were randomized into two groups: one acupuncture group and one control group, considering gender and side of hemispheral localization of lesion. With regard to the main parameters the groups were comparable at baseline. SETTING: Initially, 45 stroke patients admitted to Sunnaas Rehabilitation Hospital were included in the study: median 40 days post stroke. SUBJECTS: Forty-one of the patients were available one year after the treatment period: 21 patients in the acupuncture group and 20 controls. INTERVENTION: All subjects received an individually adapted, multidisciplinary rehabilitation programme. The acupuncture group received additional treatment with classical acupuncture for 30 min three to four times weekly for six weeks. MAIN OUTCOME MEASURES: The patients were evaluated at inclusion, after six weeks and approximately 12 months after discharge from the rehabilitation hospital. The Motor Assessment Scale (MAS) for stroke patients, Sunnaas Index of Activity of Daily Living (ADL) and Nottingham Health Profile (NHP) were used. In addition, the social situations of the patients were recorded at one year follow-up. RESULTS: The results show that the acupuncture group improved significantly more than the controls, both during the treatment period of six weeks, and even more during the following year, both according to MAS, ADL, NHP and the social situation. CONCLUSION: Although the mechanism of the effects is debatable, there seems to be a positive long-term effect of acupuncture given in the subacute stage post stroke.     

Differences in extracellular matrix proteins, epidermal growth and differentiation in discoid lupus erythematosus, lichen planus and the overlap syndrome

In this review, we focus on the heterogeneity of interstitial lung diseases detected in patients with collagen vascular diseases. By recognizing the heterogeneity of histopathology and comparing them with bronchoalveolar lavage fluid cell findings, we can understand profiles of lung inflammation and injuries and fibrosis in collagen vascular diseases. We focus on the significance of lung lymphocytosis in the lesions of patients with collagen vascular diseases, looking most closely at lesions in unusual interstitial pneumonia. The current understanding of immunopathogenesis and immunopathological findings is reviewed in the context of subsets of collagen vascular diseases.     

Normal postexercise facilitation and depression of motor evoked potentials in postpolio patients

We studied the effects of exercise on motor evoked potentials (MEPs) elicited by transcranial magnetic stimulation in healthy subjects and postpolio patients. Subjects performed repeated sets of isometric exercise until the muscle fatigued. In both groups, the mean MEP amplitude immediately after each exercise set was approximately twice that of the baseline amplitude, indicating similar postexercise facilitation, and after fatigue was approximately half that of the baseline amplitude, indicating similar postexercise depression. We conclude that the intracortical component of central fatigue is normal in postpolio patients.     

Cardiac death and stored electrograms in patients with third-generation implantable cardioverter-defibrillators

OBJECTIVE: The study's aim was to analyze the type of cardiac involvement in fetuses or infants with twin-twin transfusion syndrome. STUDY DESIGN: Seventeen pairs of monochorionic diamniotic twin fetuses with twin-twin transfusion syndrome underwent serial Doppler echocardiographic studies. Repeated decompressive amniocenteses were performed in all pregnancies. RESULTS: No specific cardiac involvement was seen in donor twins in utero or after birth. All recipient twin fetuses showed variable degrees of biventricular hypertrophy and dilation with tricuspid regurgitation. These features were also evident in 45% of the recipient twin neonates. The fluid unbalance resolved with serial amniocenteses in 9 cases and in a further 2 after the death of the donor twin; in all 11 of these recipient twin fetuses there was some degree of improvement of the cardiac involvement, which became normal in all surviving recipient twin infants within 40 days to 6 months after birth. CONCLUSION: Features of cardiac hypertrophy with signs of a prehydropic or hydropic state develop during fetal life in recipient twins in twin-twin transfusion syndrome; this impairment is reversible with the resolution of the fluid imbalance or after delivery.     

Identification of a complex congenital heart defect susceptibility locus by using DNA pooling and shared segment analysis

The isotope dilution technique of [6-3H]glucose, [U-14C]lactate and [l-14C]propionate was used to evaluate the effect of dietary chromium (Cr) supplementation on whole-body kinetics of glucose, lactate, and propionate in rams. Rams were fed a high grain diet at 2% of body weight with or without 0.5 ppm of supplemental Cr from chelated Cr for the initial 14 days, and then intake was increased to 2.5% at body weight for the last 9 days. Weight gain was enhanced (P < 0.01) with Cr supplementation. Plasma concentrations of glucose, lactate, and propionate were not influenced by Cr supplementation. Turnover rates of glucose and lactate, and their interconversion were also not influenced. Propionate turnover rate tended to increase (P = 0.11) and the conversion of propionate to glucose increased (P < 0.05) with Cr supplementation, leading the increased proportional contribution of propionate to glucose turnover rate (P < 0.05). Chromium supplementation may influence the contribution of each glucogenic substrate for glucose production in rams fed a high grain diet.     

Localization of myosin-V in the centrosome

The perinuclear localization of myosin-V was investigated in a variety of cultured mammalian cells and in primary cultures of rat hippocampus. In all cells investigated, myosin-V immunoreactivity was associated with the centrosome. In interphase cells, myosin-V was found in pericentriolar material, and in both mother and daughter centrioles. These results were obtained by using two different fixation protocols with three different affinity-purified antibodies that recognized a single band in Western blots. During cell division, myosin-V staining was intense throughout the cytoplasm and was concentrated in a trail between migrating centrioles and in the mitotic spindle poles and spindle fibers. The centrosome targeting site was determined to reside within the globular tail domain, because centrosome association also was observed in living cells transfected with DNA encoding the tail domain fused with a green fluorescent protein tag, but not in cells transfected with the vector encoding green fluorescent protein by itself.