Abstract Introduction Watch-PAT is a sleep evaluation device that measures the peripheral blood volume continuously with a probe attached to a fingertip and does not use an electroencephalogram or a nasal cannula. There has been no report on the usefulness of watch-PAT to determine the apnea diagnosis and continuous positive airway pressure (CPAP) use effects in split-night sleep study. Methods The consent of the study was obtained. Watch-PAT was simultaneously worn on a patient admitted for split-night polysomnography. The apnea-hypopnea index (AHI) obtained from PSG and the pAHI gained from the watch-PAT were measured when not using CPAP and when using CPAP respectively. And also we examined whether the reduction rates of AHI and pAHI could be correlated. Results 38 subjects (32 men, age 55 ± 13 years old). BMI 28.3 ± 5.7 kg / m2. When CPAP was not used, AHI was 57.2 ± 23.3 / h and pAHI was 50.8 ± 20.3 / h (r = 0.93, p < 0.0001), when CPAP was used, AHI was 5.2 ± 4.5 /h and pAHI was 6.2 ± 4.5 h (r = 0.82, p < 0.0001), AHI reduction rate was 90.4 ± 8.0% and pAHI reduction rate was 85.4 ± 14.6% (r = 0.76, p < 0.0001). Conclusion It was suggested that Watch-PAT had a good correlation with AHI at split night-sleep study. Support None

Transforming growth factor (TGF)-beta is known to promote tumor invasion and metastasis. Although bone morphogenetic proteins (BMPs), members of the TGF-beta family, are expressed in a variety of human carcinoma cell lines, their roles in tumor progression have not been fully clarified. In this study, we sought to determine the roles of BMPs in the progression of breast cancer bone metastasis using human breast cancer samples and a mouse xenograft model. Immunohistochemical analysis of samples from breast cancer patients as well as a mouse xenograft model of MDA-231-D, highly metastatic human breast cancer cells, revealed phospho-Smad2 and phospho-Smad1/5/8 staining in the nuclei of cancer cells in primary tumor and/or bone metastasis. Using a functional in vivo bioluminescence imaging system, we showed that TGF-beta- and BMP-induced transcriptional pathways are active in bone metastatic lesions in vivo. In addition, both TGF-beta3 and BMP-2 promoted the motility and invasiveness of the MDA-231-D cells in vitro. Moreover, expression of dominant-negative receptors for TGF-beta and/or BMPs in the MDA-231-D cells inhibited invasiveness in vitro and bone metastasis in the xenograft model. These results suggest that BMPs as well as TGF-beta promote invasion and bone metastasis of breast cancer.

Transforming growth factor (TGF)‐β signaling facilitates tumor growth and metastasis in advanced cancer. Use of inhibitors of TGF‐β signaling may thus be a novel strategy for the treatment of patients with such cancer. In this study, we synthesized and characterized a small molecule inhibitor, A‐83‐01, which is structurally similar to previously reported ALK‐5 inhibitors developed by Sawyer et al. (2003) and blocks signaling of type I serine/threonine kinase receptors for cytokines of the TGF‐β superfamily (known as activin receptor‐like kinases; ALKs). Using a TGF‐β‐responsive reporter construct in mammalian cells, we found that A‐83‐01 inhibited the transcriptional activity induced by TGF‐β type I receptor ALK‐5 and that by activin type IB receptor ALK‐4 and nodal type I receptor ALK‐7, the kinase domains of which are structurally highly related to those of ALK‐5. A‐83‐01 was found to be more potent in the inhibition of ALK5 than a previously described ALK‐5 inhibitor, SB‐431542, and also to prevent phosphorylation of Smad2/3 and the growth inhibition induced by TGF‐β. In contrast, A‐83‐01 had little or no effect on bone morphogenetic protein type I receptors, p38 mitogen‐activated protein kinase, or extracellular regulated kinase. Consistent with these findings, A‐83‐01 inhibited the epithelial‐to‐mesenchymal transition induced by TGF‐β, suggesting that A‐83–01 and related molecules may be useful for preventing the progression of advanced cancers. (Cancer Sci 2005; 96: 791–800)

Inhibitory Smads (I-Smads) repress signaling by cytokines of the transforming growth factor-beta (TGF-beta) superfamily. I-Smads have conserved carboxy-terminal Mad homology 2 (MH2) domains, whereas the amino acid sequences of their amino-terminal regions (N domains) are highly divergent from those of other Smads. Of the two different I-Smads in mammals, Smad7 inhibited signaling by both TGF-beta and bone morphogenetic proteins (BMPs), whereas Smad6 was less effective in inhibiting TGF-beta signaling. Analyses using deletion mutants and chimeras of Smad6 and Smad7 revealed that the MH2 domains were responsible for the inhibition of both TGF-beta and BMP signaling by I-Smads, but the isolated MH2 domains of Smad6 and Smad7 were less potent than the full-length Smad7 in inhibiting TGF-beta signaling. The N domains of I-Smads determined the subcellular localization of these molecules. Chimeras containing the N domain of Smad7 interacted with the TGF-beta type I receptor (TbetaR-I) more efficiently, and were more potent in repressing TGF-beta signaling, than those containing the N domain of Smad6. The isolated N domain of Smad7 physically interacted with the MH2 domain of Smad7, and enhanced the inhibitory activity of the latter through facilitating interaction with TGF-beta receptors. The N domain of Smad7 thus plays an important role in the specific inhibition of TGF-beta signaling.

Excimer laser keratectomy is widely used to correct refractive errors. Several complications of excimer laser keratectomy are reported including corneal infection, regression, corneal haze formation, glare and halo. Most of the complications are closely related to the corneal stromal wound healing process. In order to perform the excimer laser keratectomy with minimum complications, we should understand the mechanism of the corneal stroma wound healing process. In addition, such knowledge will help us to regulate the corneal stromal wound healing process in the future. In the present article, we discuss the molecular mechanism of the corneal stromal wound healing process after excimer laser keratectomy and its regulation by anti-inflammatory agents.

Abstract: TGF‐β is a potent inhibitor of cell growth, and accumulating evidence suggests that perturbation of the TGF‐β signaling pathway leads to tumorigenesis. Smads are recently identified proteins that mediate intracellular signaling of the TGF‐β superfamily. Smads 2 and 3 are phosphorylated by the TGF‐β type I receptor. Smad4 was originally identified as a candidate tumor suppressor gene in pancreatic cancers. Smads 2 and 3 form complexes with Smad4 upon TGF‐β stimulation. The heteromeric Smad complexes translocate into the nucleus, where they activate expression of target genes. Our recent study demonstrated that Smads exist as monomers in the absence of TGF‐β. Smads 2 and 3 form homo‐ as well as hetero‐oligomers with Smad4 upon ligand stimulation. Both homo‐oligomers and hetero‐oligomers directly bind to DNA, suggesting that the signaling pathway of Smads may be multiplex. Smads 2 and 3 associate with transcriptional coactivators such as p300 in a ligand‐dependent manner. p300 enhances transactivation by TGF‐β, suggesting that coactivators link Smads to the basal transcriptional machinery. A missense mutation of Smad2 identified in colorectal and lung cancers was introduced to Smad3. The mutant, Smad3(DE), blocked the activation of wild‐type Smad2 and Smad3. Thus, the missense mutation not only disrupts the function of the wild‐type Smad but also creates a dominant‐negative Smad, which could actively contribute to oncogenesis.

· Background: Transforming growth factor β (TGF-β) regulates extracellular matrix deposition, cell proliferation, and migration, and is expressed in cornea. TGF-β is thought to be involved in the corneal wound healing process. · Methods: The central corneal area (3 mm in diameter) of Lewis rats was ablated using PTK mode excimer laser and the wound healing process was observed at 12 and 24 h and 2, 5, 10, and 30 days after treatment. The expression of TGF-β1, -β2 and -β3, TGF-β type I and type II receptors, α3, α5, β4 integrin subunits, laminin and fibronectin was studied immunohistochemically. Antibody neutralizing TGF-β1, -β2 and -β3 was administered intraperitoneally, 50 µg daily, for 5 days after the laser treatment to investigate the effects of TGF-β function blockade. · Results: At the leading edge of the regenerating epithelium, no TGF-β type I and type II receptors and β4 integrin subunits were expressed after 24 h. Regenerating epithelium covered the ablated area after 2 days. An abnormal fibrotic layer was formed in the subepithelial area. This layer contained round-shaped cells in the stroma in the early stage (2–5 days after laser ablation) and spindle-shaped fibroblast-like keratocytes after 10 days. Laminin and fibronectin expression increased in the fibrotic layer. The increased stromal cells expressed TGF-β isoforms and TGF-β receptors. Neutralizing TGF-β inhibited the stromal cell increase in the laser ablated area after 5 days. · Conclusion: TGF-β may be involved in epithelial cell migration and stromal cell reaction during the corneal wound healing process after excimer laser ablation in rat models.

The transforming growth factor beta (TGF-beta) superfamily is a family of multifunctional cytokines that transduce signals via serine/threonine kinase receptors. Recent studies revealed that Mothers against dpp (Mad) in Drosophila and its homologs play important roles in the intracellular signal transduction of the serine/threonine kinase receptors. In mammals, one of the Mad homologs, MADH2 (also termed Smad2), was reported to be a mediator of TGF-beta and activin signaling and was found mutated in some of the colon and lung cancer cases. We describe here the genomic organization of the human MADH2 gene. The gene is composed of 12 exons; 2 exons 1, i.e., exon 1a and 1b, are used separately or in conjunction to form exon 1a-exon 1b-exon 2 alternatively spliced mRNA. The 2 exons 1 are closely located, and the MADH2 mRNAs are transcribed from two promoters in one CpG island. The promoter activity in the 5' upstream sequence was confirmed by the luciferase assay. The 3' end of the mRNA is heterogenous, and we found several polyadenylation signals. Northern blot analysis revealed high expression of the MADH2 mRNA, e.g., in skeletal muscle, heart, and placenta. RT-PCR assay using primers in exons 2 and 4 and direct nucleotide sequencing proved that exon 3 is spliced out in about 10% of MADH2 in human placenta. These data will be valuable for studying the MADH2 function in both normal cells and cancer cells.

Smad proteins are signal transducers for the members of the transforming growth factor‐β (TGF‐β) superfamily. Here we show that, in the absence TGF‐β stimulation, Smads exist as monomers in vivo. Smad2 and Smad3 form homo‐oligomers upon phosphorylation by the constitutively active TGF‐β type I receptor, and this oligomerization does not require Smad4. Major portions of Smad4, Smad6 and Smad7 are also present as monomers in vivo. Analysis using a cross‐linking reagent suggested that the Smad2 oligomer induced by receptor activation is a trimer. Studies by gel chromatography demonstrated that the Smad2–Smad4 heteromer is not larger than the Smad2 homomer. Moreover, overexpression of Smad4 prevented Smad2 from forming a homo‐oligomer. These findings suggest that Smad2 may form a homotrimer, or heterotrimers with Smad4, which are probably composed of two and one, or one and two molecules of Smad2 and Smad4, respectively, depending on the amount of each protein. Gel‐mobility shift assay revealed that the Smad3 homomer and Smad3–Smad4 heteromer constitute DNA‐binding complexes. Transition of the Smad proteins from monomers to oligomers is thus a critical event in the signal transduction of the TGF‐β superfamily members.

Editor,—Corneal stromal clouding (corneal haze) develops after excimer laser keratectomy. During the corneal wound healing process after excimer laser keratectomy, abnormal subepithelial fibrous tissue is formed just under the abraded area, where keratocytes proliferate and extracellular matrix (ECM) components, including collagens (type III, IV, and VII), fibronectin, laminin, and tenascin, are deposited.1-3 This abnormal ECM deposition is thought to correspond to the corneal haze. The transforming growth factor β (TGF-β) superfamily contains many multifunctional proteins, including TGF-βs, activins, and bone morphogenetic proteins (BMPs).4 They regulate cellular proliferation, differentiation, migration, and ECM production.4 Through these functions, TGF-β is known to regulate the wound healing process in many tissue. For example, TGF-β accelerates the abnormal …

Bone morphogenetic proteins (BMPs) are multifunctional cytokines, which induce bone and cartilage formation and exert various other effects on many tissues. Since angiogenesis is involved in the bone formation process, certain members in the BMP family may induce angiogenesis. We examined the in vivo angiogenic activity of BMP family members, i.e., growth/differentiation factor (GDF)-5 and BMP-2. GDF-5 induced angiogenesis in both chick chorioallantoic membrane and rabbit cornea assays. In contrast, BMP-2 did not induce angiogenesis. In order to elucidate the mechanism of angiogenesis, we examined the effects of GDF-5 on cultured bovine aortic endothelial cells (BECs). GDF-5 induced plasminogen activator activity and accelerated the migration of BECs in a chemotactic fashion, which may contribute to the process of angiogenesis in vivo. These results suggest that GDF-5 is one of the molecules which induce angiogenesis in the bone formation process.