Publications

Leptin, a hormone secreted by adipocytes, regulates the size of the adipose tissue mass through effects on satiety and energy metabolism. Leptin's precise sites of action are not known. The leptin receptor (Ob-R) is found in many tissues in several alternatively spliced forms raising the possibility that leptin exerts effects on many tissues including the hypothalamus. Ob-R is a member of the gp130 family of cytokine receptors which are known to stimulate gene transcription via activation of cytosolic STAT proteins. In order to identify the sites of leptin action in vivo, we assayed for activation of STAT proteins in mice treated with leptin. The STAT proteins bind to phosphotyrosine residues in the cytoplasmic domain of the ligand-activated receptor where they are phosphorylated. The activated STAT proteins dimerize and translocate to the nucleus where they bind DNA and activate transcription. The activation of STAT proteins in response to leptin was assayed in a variety of mouse tissues known to express Ob-R. Leptin injection activated Stat3 but no other STAT protein in the hypothalamus of ob/ob and wild-type mice but not db/db mice, mutants that lack an isoform of the leptin receptor. Leptin did not induce STAT activation in any of the other tissues tested. Activation of Stat3 by leptin was dose dependent and first observed after 15 minutes and maximal at 30 minutes. Our data indicate the hypothalamus is a direct target of leptin action and that this activation is critically dependent on the gp-130-like leptin receptor isoform missing in C57BLKS/J db/db mice. This is the first in vivo demonstration of leptin signal transduction.

The objective of this study was to identify risk factors for work disability among persons with carpal tunnel syndrome (CTS). The study was designed to analyze data from the Occupational Health Supplement of the National Health Interview Survey, a nationwide, population-based survey. Subjects included 544 survey respondents with self-report of CTS and 32,688 survey respondents without CTS, all aged 18-64 years, and with a history of labor force participation. Measurements were as follows: Dependent variables were work disability, defined either as cessation of employment without attribution of cause or, alternatively, as cessation of employment or job change specifically attributed to CTS by the survey respondent. Independent variables were ergonomic risk of work disability, defined by minutes of workplace repetitive hand and wrist bending for the most recent job held. This measure was derived from responses categorized by an occupation and industry matrix independent of CTS status. Socio-demographic and health status risk factors for work disability were based on the respondent report. The main results were as follows: Among 544 persons with CTS, 58 (11%, CI 8-13%) reported work disability specifically attributed to CTS, representing an estimated national prevalence of 240,578 persons with this limitation. Workplace ergonomic risk, measured as repetitive hand or wrist bending in the occupation and industry of last employment, was a significant factor predictive of CTS-attributed work disability (per 120 min of daily exposure, OR 1.7, CI 1.1-2.6), even after taking into account socio-demographic factors and health status. The conclusions were that work disability among persons with CTS is common. For those with CTS, working conditions characterized by repetitive bending of the hand or wrist may increase the risk of work disability associated with this condition.

To examine whether diffuse pleural thickening (DPT) causes impairment of pulmonary function independent of other manifestations of asbestos-related disease, we studied individuals selected from 1,150 men with occupational asbestos exposure who had undergone pulmonary function testing and computed tomographic (CT) scanning. The CT scans revealed 84 subjects with DPT as defined for CT. Of these 84 subjects, 53 eligible study cases were matched by age with a referent group without DPT from the same exposed group. No difference was demonstrated between cases and referents in smoking history, length of exposure, latency, or the proportion with either interstitial fibrosis or pleural plaques. Individuals with DPT demonstrated significantly reduced forced vital capacity (FVC) (p = 0.002) and diffusing capacity for carbon monoxide (DLCO) (p = 0.002) as compared with the referents. No difference was found in the two groups' FEV1 to FVC ratio (FEV1/FVC). Individuals with DPT and either interstitial fibrosis or pleural plaques showed a significantly lower FVC than did those with fibrosis or pleural plaques alone. Individuals with DPT and rounded atelectasis had similar pulmonary function to those with DPT and no rounded atelectasis. Subjects with DPT had a more frequent history of coronary bypass surgery than referents (19% versus 2%; p = 0.008). We conclude that subjects with DPT have restrictive pulmonary function and reduced diffusing capacity independent of other manifestations of asbestos-related disease.

The hepatic hemoprotein tryptophan 2,3-dioxygenase (TDO) is the key regulatory enzyme that, through irreversible degradation, controls the flux of tryptophan through physiologically relevant pathways. This enzyme is composed of four identical subunits and in its fully assembled tetrameric form requires 2 mol of heme (Fe(+2)-protoporphyrin IX)/mol of protein for functional competence. Using a full-length cDNA for the rat liver TDO subunit (pUC119/TDO) as the template, TDO cDNA was amplified by polymerase chain reaction (PCR) and incorporated into the expression vector pTrc99A after introduction of convenient restriction sites as well as modification of the second codon AGT to GCT to optimize its bacterial expression. DH5 alpha F' strain Escherichia coli cells transfected with this pTrc99A/TDO construct expressed soluble, functionally active, tetrameric TDO protein in high yields. The enzyme was isolated from 30,000g supernatant of cell lysates, purified by ion-exchange chromatography, and its spectral and catalytic properties were assessed in terms of its substrate and prosthetic moiety specificities. In almost all aspects, the bacterially expressed enzyme was found to be identical to that of the rat liver. Heterologous expression of the fully functional enzyme, we trust, will enable future elucidation of its structure-function relationships.