Retroviral display of antibody fragments; interdomain spacing strongly influences vector infectivity

Five different single-chain antibody fragments (scFv) against human cell-surface antigens were displayed on murine ecotropic retroviral vectors by fusing them to the Moloney SU envelope glycoprotein. The spacing between the scFv and the SU glycoprotein was varied by fusing the scFv to residue +7 or to residue +1 of Moloney SU and by inserting linker sequences of different lengths between the domains. All of the chimeric envelopes were efficiently incorporated into vector particles and could bind to human cells through their displayed antibody fragments, but did not infect them. The spacing between the scFvs and the SU glycoproteins had no significant effect on the efficiency of envelope expression or viral incorporation and did not affect the binding properties of the chimeric envelopes, nor did it influence the efficiency of targeted gene delivery to human cells by scFv-displaying vectors. However, on murine fibroblasts the infectivity of vectors incorporating the chimeric envelopes was strongly influenced by the length of the interdomain spacer. The titers were very low when the single-chain antibodies were fused through a tripeptide linker to SU residue +7 and were greatly enhanced (up to 10(5)-fold) when they were fused to SU residue +1 through a heptapeptide linker. These results point to the importance of steric interactions between the domains of chimeric envelope glycoproteins and may have implications for retroviral vector design for human gene therapy.     

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.     

Cell-type-specific expression of rat steroid 5 alpha-reductase isozymes

The enzyme steroid 5 alpha-reductase (EC 1.3.99.5) is a component of an intercellular signaling pathway that determines cell fate in the primordium of the mammalian reproductive tract. During male phenotypic sexual differentiation, the dihydrotestosterone product of this enzyme binds to the androgen receptor and initiates development of the external genitalia and prostate. Genes encoding two isozymes of steroid 5 alpha-reductase with different biochemical properties and tissue distributions have recently been isolated. In the current study, we utilize in situ hybridization analysis to determine cell-type-specific expression patterns of the 5 alpha-reductase isozyme mRNAs in two androgen target tissues (regenerating ventral prostate and epididymis) and a peripheral tissue (liver). In regenerating ventral prostate, the type 1 mRNA is expressed in basal epithelial cells whereas expression of the type 2 mRNA is largely confined to stromal cells. These results were confirmed by immunohistochemical analysis and are consistent with distinct roles played by the isozymes in the prostate. In the epididymis, both 5 alpha-reductase isozyme mRNAs are expressed in epithelial cells. Only the type 1 mRNA is present in the liver. This mRNA is distributed in a striking spatial gradient extending from hepatocytes surrounding the portal triad (high expression) to those surrounding the central vein (low to absent expression). These findings demonstrate cell-type-specific expression of the steroid 5 alpha-reductase isozymes and underscore their distinct and overlapping functions in androgen physiology.     

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.     

Antecedents and outcomes of work-family conflict: Testing a model of the work-family interface.

A comprehensive model of the work–family interface was developed and tested. The proposed model extended prior research by explicitly distinguishing between work interfering with family and family interfering with work. This distinction allowed testing of hypotheses concerning the unique antecedents and outcomes of both forms of work–family conflict and a reciprocal relationship between them. The influence of gender, race, and job type on the generalizability of the model was also examined. Data were obtained through household interviews with a random sample of 631 individuals. The model was tested with structural equation modeling techniques. Results were strongly supportive. In addition, although the model was invariant across gender and race, there were differences across blue- and white-collar workers. Implications for future research on the work–family interface are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)