Publications

Human asthma is characterized by increased airway hyperreactivity to a variety of bronchoconstricting agents. Aberrant type 2 immune responses in the lung have been associated with airway hyperreactivity in both human asthma and in murine models of allergic airways disease. Despite their intrinsically elevated basal airway reactivity to smooth muscle constricting agents, A/J mice demonstrated no inherent inflammatory cell infiltration nor elevation of type 2 cytokines in the lung. Crossed bone marrow reconstitution experiments between A/J and MHC congenic B10.A mice revealed enhanced airway reactivity only in A/J recipients, irrespective of whether they had been reconstituted with A/J or B10. A hemopoietic cells. Further, A/J-derived bone marrow cells did not affect the reactivity of B10.A recipients. Although mice on RAG-deficient and IL-4-deficient backgrounds demonstrate substantial abrogation of allergen-induced airway hyperreactivity, these gene deletions had no impact on the elevated baseline reactivity when backcrossed onto A/J mice. Thus, in these mice, basal airway hyperreactivity is maintained independently of type 2 immunity induced by allergens.

The integrin alpha9beta1 mediates neutrophil migration across several ligands that are enriched at sites of inflammation. In one of these ligands, the acidic phosphoprotein osteopontin, the alpha9beta1 binding site is cryptic, but is revealed after thrombin cleavage. We have recently mapped the alpha9beta1 binding site in osteopontin to the linear peptide sequence, SVVYGLR, immediately adjacent to the thrombin cleavage site. Interestingly, this site is also adjacent to a sequence (RGD) through which five other integrins bind to osteopontin. These findings suggest a novel mechanism by which thrombin can modulate integrin signaling at sites of tissue injury.

The hepatic cytosolic hemoprotein tryptophan 2,3-dioxygenase (TDO) is the rate-limiting enzyme in tryptophan catabolism and thus plays a key role in regulating the physiological flux of tryptophan into relevant metabolic pathways. The TDO protein is induced by corticosteroids such as dexamethasone (DEX) and is stabilized by its prosthetic heme. In rats, acute chemically induced hepatic heme depletion reduces the functional hepatic TDO levels to 25-30% of basal levels within 1 h, and this decrease persists beyond 28 h of heme depletion at which time only 25-30% of the protein is available for heme incorporation. Since this could stem from impaired de novo synthesis and/or instability of the newly synthesized apoTDO protein in the absence of heme, we examined the specific role of heme in these events in a previously validated rat model of acute hepatic heme depletion triggered by the P450 suicide substrate 3, 5-dicarbethoxy 2,6-dimethyl-4-ethyl-1,4-dihydropyridine. We now show that exogenous heme can reverse the functional impairment of the enzyme observed during hepatic heme depletion and fully restore the impaired DEX-mediated induction of the enzyme to normal. Furthermore, through Northern/slot blot analyses coupled with nuclear run-on studies, we now document that this heme regulation of TDO is exerted primarily at the transcriptional level. Immunoblotting analyses also reveal corresponding changes in the TDO protein, thereby establishing that heme is necessary for DEX-inducible TDO mRNA transcription and subsequent translation. Thus, the TDO gene may contain heme-regulatory elements in addition to the reported glucocorticoid-responsive elements. Together, these findings suggest that clinically, hepatic heme deficiency may enhance the tryptophan flux into synthetic (serotonergic) pathways, not only by depriving prosthetic heme for a functionally competent TDO hemoprotein, its primary catabolic enzyme, but also by impairing the de novo synthesis of this enzyme.

Cardiomyopathy is a major cause of morbidity and mortality. Ventricular conduction delay, as shown by prolonged deflections in the electrocardiogram caused by delayed ventricular contraction (wide QRS complex), is a common feature of cardiomyopathy and is associated with a poor prognosis. Although the G(i)-signaling pathway is up-regulated in certain cardiomyopathies, previous studies suggested this up-regulation was compensatory rather than a potential cause of the disease. Using the tetracycline transactivator system and a modified G(i)-coupled receptor (Ro1), we provide evidence that increased G(i) signaling in mice can result in a lethal cardiomyopathy associated with a wide QRS complex arrhythmia. Induced expression of Ro1 in adult mice resulted in a >90% mortality rate at 16 wk, whereas suppression of Ro1 expression after 8 wk protected mice from further mortality and allowed partial improvement in systolic function. Results of DNA-array analysis of over 6,000 genes from hearts expressing Ro1 are consistent with hyperactive G(i) signaling. DNA-array analysis also identified known markers of cardiomyopathy and hundreds of previously unknown potential diagnostic markers and therapeutic targets for this syndrome. Our system allows cardiomyopathy to be induced and reversed in adult mice, providing an unprecedented opportunity to dissect the role of G(i) signaling in causing cardiac pathology.

BACKGROUND

The ability to predict the rate of hemodynamic progression in an individual patient with valvular aortic stenosis has been elusive. The purpose of the present study was to evaluate whether the rate of change in aortic valve area (AVA) measured during the ejection phase of a cardiac cycle predicts the rate of hemodynamic progression in patients with asymptomatic aortic stenosis.

METHODS AND RESULTS

In 84 adults with initially asymptomatic aortic stenosis and a baseline AVA of > or =0.9 cm(2), annual echocardiographic data were obtained prospectively (mean follow-up 2.8+/-1.3 years). With the initial echocardiogram, the ratio of AVA measured at mid-acceleration and mid-deceleration to the AVA at peak velocity was calculated. The primary outcome variable was the annual rate of change in AVA (rate of progression), with rate of progression classified as rapid (a reduction in AVA of > or =0.2 cm(2)/y) or slow (<0.2 cm(2)/y). Rapid progression was significantly associated with an AVA ratio of > or =1.25 (P=0.004, risk ratio 3.1, 95% CI 1.2 to 7.9). The sensitivity, specificity, and positive predictive value of AVA ratio of > or =1.25 for the prediction o rapid progression of valvar aortic stenosis was 64%, 72%, and 80% respectively. The decrease in ejection fraction measured from the initial to final echocardiogram was small but greater for patients with an AVA ratio of > or =1.25 (-4+/-7% versus +2+/-7%, P<0.001).

CONCLUSIONS

A flow-dependent change in AVA can be measured during a routine transthoracic echocardiographic study. The rate of change in AVA is an additional measure of disease severity and may be used to predict an individual's risk for subsequent rapid disease progression.

BACKGROUND

Although the cure of breast cancer by "early detection" and prompt treatment rests on the belief that all breast cancers grow at the same rate, many cancers have been shown to grow rapidly and others slowly. In particular, mammography screening may often detect the slow-growing, nonaggressive tumors that might not be found until much later, if at all.

METHODS

We reviewed the medical records of a natural cohort of 233 patients. The cohort comprised all women who received their first antineoplastic treatment for breast cancer at Yale-New Haven Hospital during the period from January 1 through December 31, 1988, and had a median follow-up thereafter of 82.4 months.

RESULTS

The mammography screen-detected group (MSDG) contained 97 (42%) of the 233 breast cancers. The rates of subsequent freedom from cancer deaths or recurrences were 95% (92 patients) in the MSDG and 79% (107 patients) in all other patients (log-rank 2P<.001). This superiority occurred partly because 90 (93%) of the MSDG were in the good prognosis TNM stages 0, I, and IIA, compared with 92 (68%) of the non-MSDG (chi2 2P = .001). Of the 31 patients with stage 0 (carcinoma in situ), all of whom had disease-free survival, 24 (77%) were found by mammography screening. Even within similar TNM stages, however, the MSDG had distinctly better disease-free survival results than the non-MSDG. For patients in TNM stages I and IIA, the "failure events" had respective rates of 2% and 13% (log-rank 2P = .02).

CONCLUSIONS

The results suggest that many of the breast cancers found by mammography screening have excellent prognosis not just because of early detection, but also because many of the cancers are relatively benign, requiring minimal therapy.