HER-2/c-erbB2 is phosphorylated by calmodulin-dependent protein kinase II on a single site in the cytoplasmic tail at threonine-1172

Calmodulin-dependent protein kinase II (Cam kinase II) is known to desensitise epidermal growth factor receptor (HER-1) tyrosine kinase activity by a process involving phosphorylation at serines 1046/47 in the cytoplasmic tail. We have developed an experimental system to investigate phosphorylation of the related HER-2/c-erbB2 proto-oncogene utilising purified Cam kinase II and recombinant glutathione-S-transferase (GST) fusion proteins. The cDNA for rat Cam kinase II-alpha was transfected into human embryonic kidney (HEK) 293 fibroblasts and the expressed protein purified to homogeneity by calmodulin-agarose affinity chromatography. A GST fusion protein comprising residues 1126-1255 of HER-2 was phosphorylated by purified Cam kinase II, in contrast to a GST protein comprising residues 1005-1125. Phosphoamino-acid analysis and site-directed mutagenesis indicated that HER-2 was phosphorylated on a single site at threonine-1172 which resides within a consensus Cam kinase II phosphorylation site (RAKT). HER-2 (threonine-1172-alanine), in the form of a ligand-inducible chimaera HER-1/2, was co-transfected into HEK-293 fibroblasts with a constitutively active form of Cam kinase II, followed by in vivo labelling of these cells with 32 P-orthophosphate. Immunoprecipitation of ligand-activated receptors followed by two-dimensional phosphopeptide mapping indicated that threonine-1172 in HER-2 is a newly identified in vivo site which can be hyper-phosphorylated by constitutively active Cam kinase II. In addition, when over-expressed in HEK-293 fibroblasts, HER-1/2 (threonine-1172-alanine) showed a defect in desensitisation and underwent a more sustained EGF-induced receptor autophosphorylation compared to wild-type HER-1/2.     

A monitoring, feedback, and triggering system for reproducible breath-hold MR imaging

OBJECTIVE: To evaluate the diagnostic accuracy and clinical usefulness of high-resolution transvaginal duplex Doppler ultrasound in postpartum and post-abortion patients with excessive hemorrhage who are suspected of having residual trophoblast. METHODS: Forty-eight women with excessive hemorrhage referred for possible residual trophoblastic tissue were evaluated by transvaginal duplex Doppler ultrasonography. Based on two-dimensional imaging, the patients were divided prospectively into groups: women who had an empty uterus with a normal uterine cavity, those with a pure endometrial fluid collection and no echogenic foci, those who had a mixed endometrial fluid collection with foci of echogenicity, and those with intracavitary heterogeneous material with mixed echo patterns of fluid and solid components. In each group, Doppler studies were performed and the resistance index (RI) was calculated. The two-dimensional patterns and Doppler results were correlated with clinical and pathologic follow-up. RESULTS: Twenty-eight subjects had a normal uterine cavity and seven had a pure endometrial fluid collection; all were treated conservatively and none showed later clinical evidence of residual trophoblastic tissue. In 13 women, residual trophoblast was strongly suggested from the images of two-dimensional ultrasonography: Five showed an endometrial fluid collection with some echogenic foci, and eight exhibited intracavitary mixed echogenic material. All underwent curettage, and residual trophoblastic tissue was found in ten of the 13. The mean (+/- standard deviation) RI to flow in the myometrial arteries was 0.54 +/- 0.15 in women without residual trophoblast and 0.35 +/- 0.1 in those with residual trophoblastic tissue (P < .01). CONCLUSION: Our experience suggests that transvaginal duplex Doppler ultrasonography is an effective noninvasive method for evaluating patients with excessive postpartum and post-abortion hemorrhage who are suspected of having residual trophoblastic tissue. Its use enhances the positive preoperative diagnosis of residual trophoblastic tissue and may reduce unnecessary curettage procedures.     

Characterization of type I 5 alpha-reductase activity in DU145 human prostatic adenocarcinoma cells

Each amino acid is represented by a vector of numerical measurements for the attributes of volume, area, hydrophilicity, polarity, hydrogen bonding, shape, and charge. Inter-residue distances are then calculated according to common metrics, and we introduce a new clustering objective function derived from information-theoretic considerations. The arguments of the function are the inter-object distances of the things to be clustered: in this case the amino acids. By means of approximating the solution of an integer programming problem, then, the residues are partitioned into clusters. The clusters obtained are compared with groups obtained in substitution/mutation studies and found to be similar. Thus, probably the strongest and most objective evidence to date is supplied for believing that physico-chemical properties account for the viability of substitutions and that the important similarities/differences are explained by a relatively small and simple set of properties.     

Measurement error and results from analytic epidemiology: dietary fat and breast cancer

BACKGROUND: International correlational analyses have suggested a strong positive association between fat consumption and breast cancer incidence, especially among post-menopausal women. However, case-control studies have been taken to indicate a weaker association, and a recent, pooled cohort analysis reported little evidence of an association. Differences among study results could be due to differences in the populations studied, differences in the control for total energy intake, recall bias in the case-control studies, and dietary measurement error biases. Existing measurement error models assume either that the sample data used to validate dietary self-report instruments are without measurements error or that any such error is independent of both the true dietary exposure and other study subject characteristics. However, growing evidence indicates that total energy and, presumably, both total fat and percent energy from fat are increasingly underreported as percent body fat increases. PURPOSE: A relaxed dietary measurement model is introduced that allows all measurement error parameters to depend on body mass index (weight in kilograms divided by the square of height in meters) and incorporates a random underreporting quantity that applies to each dietary self-report instrument. The model was applied to results from international correlational analyses to determine whether the differing associations between dietary fat and postmenopausal breast cancer can be explained by measurement errors in dietary assessment. METHODS: The relaxed measurement model was developed by use of data on total fat intake and percent energy from fat from 4-day food records (4DFRs) and food-frequency questionnaires (FFQs) from the original Women's Health Trial. This trial was a randomized, controlled, feasibility study of a low-fat dietary intervention carried out from 1985 through 1988 in Cincinnati (OH), Houston (TX), and Seattle (WA) among 303 women (184 intervention and 119 control) who were 45-69 years of age. The relaxed model was used to project results from the international correlational analyses onto 4DFR and FFQ fat-intake categories. RESULTS AND CONCLUSIONS: If measurement errors in dietary assessment are overlooked entirely, the projected relative risks (RRs) for breast cancer based on the international data vary substantially across percentiles of total fat intake. The projected RR for the 90% versus the 10% fat-intake percentile is 3.08 with the 4DFR and 4.00 with the FFQ. If random (i.e., noise) aspects of measurement error are acknowledged, the projected RR for the same comparison is reduced to 1.54 with the 4DFR and 1.42 with the FFQ. If both systematic and noise aspects of measurement error are acknowledged, the projected RR is reduced to about 1.10 with either instrument. Acknowledgment of measurement error also leads to a projected RR of about 1.10 for the 90% versus the 10% percentile of percent energy from fat with either dietary instrument. IMPLICATIONS: Dietary self-report instruments may be inadequate for analytic epidemiologic studies of dietary fat and disease risk because of measurement error biases.     

The importance of geriatrics to family medicine: a position paper by the Group on Geriatric Education of the Society of Teachers of Family Medicine

The role of geriatrics and geriatricians in family medicine remains unsettled. Despite a rapidly aging population, a tremendous shortage now exists of faculty with interest and expertise in geriatrics. Relatively few family practice residents choose to enter geriatric fellowship programs, and federal funding for such programs has been reduced. Despite accreditation requirements, residency programs are not always able to provide the range of geriatric experiences needed to properly prepare graduates to provide care for the broad range of older patients. Medical students' exposure to geriatrics remains limited. The Group on Geriatric Education of the Society of Teachers of Family Medicine believes that family medicine faculty must recognize and be committed to the notion that geriatrics is integral to family medicine. Both undergraduate and residency training programs should emphasize experience with geriatric patients in multiple settings. In particular, the nursing home should not be the main focus of geriatric training. The small number of certified geriatric faculty will be able to provide leadership, but a broad range of faculty must become involved in teaching geriatrics. Faculty development activities and continuing education programs to foster the necessary expertise will be essential to the accomplishment of this task.