Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertension

BACKGROUND: Chronic hypoxia influences gene expression in the lung resulting in pulmonary hypertension and vascular remodelling. For specific investigation of the vascular compartment, laser-microdissection of intrapulmonary arteries was combined with array profiling. METHODS AND RESULTS: Analysis was performed on mice subjected to 1, 7 and 21 days of hypoxia (FiO(2 )= 0.1) using nylon filters (1176 spots). Changes in the expression of 29, 38, and 42 genes were observed at day 1, 7, and 21, respectively. Genes were grouped into 5 different classes based on their time course of response. Gene regulation obtained by array analysis was confirmed by real-time PCR. Additionally, the expression of the growth mediators PDGF-B, TGF-β, TSP-1, SRF, FGF-2, TIE-2 receptor, and VEGF-R1 were determined by real-time PCR. At day 1, transcription modulators and ion-related proteins were predominantly regulated. However, at day 7 and 21 differential expression of matrix producing and degrading genes was observed, indicating ongoing structural alterations. Among the 21 genes upregulated at day 1, 15 genes were identified carrying potential hypoxia response elements (HREs) for hypoxia-induced transcription factors. Three differentially expressed genes (S100A4, CD36 and FKBP1a) were examined by immunohistochemistry confirming the regulation on protein level. While FKBP1a was restricted to the vessel adventitia, S100A4 and CD36 were localised in the vascular tunica media. CONCLUSION: Laser-microdissection and array profiling has revealed several new genes involved in lung vascular remodelling in response to hypoxia. Immunohistochemistry confirmed regulation of three proteins and specified their localisation in vascular smooth muscle cells and fibroblasts indicating involvement of different cells types in the remodelling process. The approach allows deeper insight into hypoxic regulatory pathways specifically in the vascular compartment of this complex organ

Hospitalization for ambulatory care-sensitive conditions: a method for comparative access and quality studies using routinely collected statistics.

BACKGROUND: Appropriate and timely provision of ambulatory care is an important factor in maintaining population health and in avoiding unneccessary hospital use. This article describes conditions for which hospitalization rates have a strong and inverse relationship to access to high-quality ambulatory care. METHODS: Three panels of Canadian physicians following different consensus techniques selected conditions for which the relative risk of hospitalization is inversely related to ambulatory care access. PRINCIPAL FINDINGS: All panels identified asthma, angina pectoris, congestive heart failure, otitis media, gastric ulcer, pelvic inflammatory disease, malignant hypertension, and immunization-preventable infections as ambulatory care-sensitive admissions. These conditions strongly overlap with lists developed for similar purposes in the U.S. and England. INTERPRETATION: Ambulatory care-sensitive conditions represent an intermediate health outcome. They are distinct from inappropriate hospitalizations. They may be useful for measuring the impact of health care policy, and for performance measurement or audit