Drug Binding to α1-Acid Glycoprotein Studied by Circular Dichroism

The interactions of acidic and basic drugs with ₁-acid glycoprotein (₁-AGP) were investigated using circular dichroism (CD) measurements. Extrinsic Cotton effects were generated by the binding of drugs to ₁-AGP. The CD data suggested the presence of a single binding site on the ₁-AGP molecule. The induced ellipticities of the acidic drug–₁-AGP system decreased with increasing pH, while the ellipticities for the basic drugs increased with pH. The ellipticities for all drugs were reduced by the addition of fatty acids. Furthermore, the induced ellipticities decreased in the presence of cesium chloride for basic drugs bound to ₁-AGP. The extrinsic Cotton effects therefore appear to result from hydrophobic interaction with ₁-AGP for the acidic drugs and from hydrophobic and electrostatic interactions for the basic drugs.     

Directed differentiation of telencephalic precursors from embryonic stem cells

We demonstrate directed differentiation of telencephalic precursors from mouse embryonic stem (ES) cells using optimized serum-free suspension culture (SFEB culture). Treatment with Wnt and Nodal antagonists (Dkk1 and LeftyA) during the first 5 d of SFEB culture causes nearly selective neural differentiation in ES cells ( approximately 90%). In the presence of Dkk1, with or without LeftyA, SFEB induces efficient generation ( approximately 35%) of cells expressing telencephalic marker Bf1. Wnt3a treatment during the late culture period increases the pallial telencephalic population (Pax6(+) cells yield up to 75% of Bf1(+) cells), whereas Shh promotes basal telencephalic differentiation (into Nkx2.1(+) and/or Islet1/2(+) cells) at the cost of pallial telencephalic differentiation. Thus, in the absence of caudalizing signals, floating aggregates of ES cells generate naive telencephalic precursors that acquire subregional identities by responding to extracellular patterning signals.     

Epidemiological study on association of periodontal disease and total and differential leukocyte counts

Periodontal disease is defined as inflammation that is caused by bacteria in dental plaque. This disease is liable to be a factor contributing to the high leukocyte count over an extended period. Furthermore, a number of prospective epidemiologic studies have shown that the leukocyte count is a good predictor of ischemic heart disease (IHD). However there have been few epidemiological studies of the relationship between periodontal disease and the leukocyte count. The purpose of this study was to investigate the relationship between the total and differential leukocyte counts and oral conditions of Japanese factory workers who were classified according to their smoking habits. The 1,167 subjects were male factory workers employed with a chemical factory in Osaka, Japan. The oral conditions recorded were periodontal status (Community Periodontal Index of Treatment Needs, CPITN). The relationship between the total and differential leukocyte counts and the CPITN score of subjects who were classified according to their smoking habits was investigated in 1996 and 1997. In both the current smokers and nonsmokers, the subjects with severe periodontal disease, in contrast to the normal subjects, exhibited total leukocyte, neutrophil and monocyte counts that were significantly high for 1 year during follow-up studies. The periodontal disease is shown that causes the total leukocyte and neutrophil counts related to the development of IHD to remain at a high level.     

Cohort study of the total leukocyte count and periodontal disease among company employees

There have been few epidemiological studies of the relationship between the leukocyte count and dental disease. In the present study, therefore, we investigated the relationship between oral health indicated by the Community Periodontal Index of Treatment Needs (CPITN) and the total leukocyte count in the cohort study. The 1,035 subjects were male factory workers employed at a chemical factory in Osaka, Japan. Their oral conditions were recorded as the CPITN score. The relationship between the total leukocyte count and the oral condition of the subjects classified according to their smoking habits was investigated over a 5-year period. Among the current smokers, the total leukocyte count was highest each year for the group with CPITN level 4, followed by those with CPITN levels 3 and 2 in descending order, showing that the total leukocyte count was reduced as the periodontal diseases ameliorated. Among the nonsmokers, the total leukocyte count was high every year in the group with CPITN levels 4 or 3 compared to that of the group with CPITN level 2. The fluctuations of the total leukocyte count in current smokers and nonsmokers whose CPITN score increase or decreased in five years was investigated. There were no significant differences among the all groups.     

Photoreactive nanotool for cell aggregation and immobilization

We developed a new method to prepare aggregates of specific cells and immobilize cells on a substrate with specific shapes by using a synthetic multifunctional tool, which consisted of a cell adhesive Arg-Gly-Asp (RGD) sequence, a photoreactive phenyl azido group and a biotin group. This chemical nanotool, RGD-2-(6-[biotinamido]-2-(p-azidobenzamido)-hexaneamido)ethyl-1,3′-dithio-proprionate (RGD-BED) was added to human umbilical vein endothelial cells to bind to receptors via the ligand–receptor interaction. Next a photoimmobilization of the binding RGD-BED was carried out by UV irradiation to covalently couple a phenyl azido moiety of RGD-BED with the neighboring site of the integrin receptor. We found that not only the migration distance of RGD-BED immobilized cells was diminished, but also the cell morphology was fixed on the substrate due to the blocking of integrin receptors by RGD-BED. In contrast, the addition of biocytin-containing polymers, poly(N-methacryloyloxy biocytin-co-dimethylacrylamide), and avidin to the RGD-BED-immobilized cells led to restore cellular migration behavior, probably arising from the increase in the rigidity of the environment surrounding the cells. Furthermore, by the addition of avidin to the RGD-BED-immobilized cells, three-dimensional cell aggregates were formed due to the cross-linking of the biotin moieties of RGD-BED. These results show that RGD-BED is a potential nanotool not only to label and collect targeted cells by the formation of cell aggregates but also to suppress mobility and morphologies of specific cells to be applicable for medical treatments.     

Tranilast inhibits the proliferation of uterine leiomyoma cells in vitro through G1 arrest associated with the induction of p21(waf1) and p53

Uterine leiomyoma is a mesenchymal tumor composed of smooth muscle cells with fibrous tissues and many mast cells. Tranilast is known to suppress fibrosis or to work as a mast cell stabilizer and is reported to inhibit proliferation of vascular smooth muscle cells. In this study, we examined the effects of tranilast on cultured human leiomyoma cells in vitro to evaluate whether this agent has the potential to inhibit the growth of uterine leiomyomas. Tranilast inhibited the proliferation of cultured leiomyoma cells in a dose-dependent manner without any cytotoxic effect or induction of apoptosis. In association with the inhibitory effect, tranilast induced the cyclin-dependent kinase (CDK) inhibitor p21(waf1) and tumor suppressor gene p53 and decreased CDK2 activity. These results suggest that tranilast arrests the proliferation of uterine leiomyoma cells at the G0/G1 phase, through the suppression of CDK2 activity via an induction of p21(waf1) and p53. Tranilast was concluded to be a potent agent to inhibit proliferative activity of uterine leiomyoma cells.     

Role of electrophysiologic study (EPS)-guided preventive therapy for the management of ventricular tachyarrhythmias in patients with heart failure.

BACKGROUND: Ventricular tachyarrhythmias (VT/VF) are 1 of the most important factors determining the prognosis of patients with heart failure (HF). Although priority is given to implantable cardioverter defibrillator (ICD) therapy for the prevention of sudden cardiac death, electrophysiologic-study (EPS)-guided preventive therapy could be important for reducing the number of cardiac events. METHODS AND RESULTS: Of 864 patients with a history of HF, an EPS was performed in 168 and 121 had inducible VT/VF. Under the basic therapy of an ICD, additional catheter ablation was attempted for 95 of 124 monomorphic VT foci in 74 patients, and 78 of the VT were successfully ablated. The prognoses were compared among 5 patient groups with different results for the EPS and catheter ablation: (1) success group (n=43), (2) failure group (n=15), (3) not attempted group (n=16), (4) VF group (n=47), and (5) no inducible VT/VF group. During a follow-up period of 31+/-22 months, the incidence of VT/VF was lower in the success and no inducible VT/VF groups than in the other groups (p=0.0018), although a significant difference was not observed for the total deaths. CONCLUSION: EPS-guided preventive therapy using an ICD and catheter ablation can be useful, at least for the reduction of arrhythmic events in patients with HF.