Bacteriophage T3 DNA binding protein interaction with DNA

Single-stranded DNA binding protein (DBP), coded by T3 phage, is essential for concatemer formation (H. Fujisawa, M. Yamagishi, H. Matsuo-Kato, and T. Minagawa, 1980, Virology, 105, 480–489.) T3 DBP was purified to homogeneity and found to exist as a dimer. The structure of the T3 DBP-fd DNA complex appears as a condensed and beaded ring structure by electron microscopy. As judged by sucrose gradient centrifugation, DBP binds to single-stranded, but not to double-stranded DNA; at saturation, one protein monomer is bound per every 100 nucleotides. The T3 DBP has a strong ability to catalyze renaturation of DNA even in the absence of Mg. The results suggest that the T3 DBP acts in concatemer formation by stimulating pairing of the single-stranded redundant termini of T3 DNA.     

New selection criterion for Fontan procedure: pulmonary artery clamping test and pulmonary vascular resistance in increased pulmonary blood flow

A new selection criterion for Fontan procedure, pulmonary artery clamping test (PACT) was developed and employed in 13 candidates for Fontan procedure. PACT was aimed to evaluate the response of pulmonary vascular bed to increased pulmonary blood flow and calculate the pulmonary vascular resistance in the increased pulmonary blood flow as a selection criterion for Fontan procedure, preoperatively. After a median sternotomy incision, an electromagnetic flow meter was employed for measuring the pulmonary blood flow (Qp) on the pulmonary trunk. By clamping the left pulmonary artery and the right pulmonary artery, right pulmonary vascular resistance and the left pulmonary resistance in the pulmonary blood flow of Qp were calculated respectively. By means of electrical analogue, right and left lungs were simulated as resistors in parallel. And the equivalent pulmonary vascular resistance Rpc was calculated as the predicted pulmonary vascular resistance in the increased pulmonary blood flow of two times Qp. Fontan procedure was performed in 9 cases with a Rpc of less than 3 unit.m2 with a successful result and no death. Postoperative cardiac index ranged from 2.6 to 3.7 L/min.m2 with a mean of 2.8 L/min.m2. Postoperative pulmonary vascular resistance ranged from 1.6 to 3.2 unit.m2. Postoperative cardiac index was correlated with postoperative pulmonary vascular resistance. And postoperative pulmonary vascular resistance was well correlated with equivalent pulmonary vascular resistance but was not correlated with preoperative pulmonary vascular resistance measured by Fick's method at the preoperative catheterization. In Fontan procedure, the pulmonary blood flow frequently increases to a certain degree postoperatively. And pulmonary vascular resistance is not a static resistance, but a dynamic resistance. Hence, in discussing a pulmonary vascular resistance as a selection criterion for Fontan procedure, the pulmonary blood flow in which the pulmonary vascular resistance stand should be taken into account. Equivalent pulmonary vascular resistance Qpc is a theoretical selection criterion for Fontan procedure and well reflects the postoperative pulmonary vascular resistance. Fontan procedure can be successfully performed with a Rpc of less than 3 unit.m2.     

Chromosomal aberrations in human hepatocellular carcinomas associated with hepatitis C virus infection detected by comparative genomic hybridization.

Thirty-five hepatocellular carcinomas (HCCs) associated with hepatitis C virus (HCV) were analysed by comparative genomic hybridization (CGH), to screen for changes in copy-number of DNA sequences. Chromosomal losses were noted in 1p34-36 (37%), 4q12-21 (48%), 5q13-21 (35%), 6q13-16 (23%), 8p21-23 (28%), 13q (20%), 16q (33%) and 17p13 (37%). Gains were noted in 1q (46%), 6p (20%), 8q21-24 (31%) and 17q (43%). High level gains indicative of gene amplifications were found in 7q31 (3%), 11q13 (3%), 14q12 (6%) and 17q12 (3%); amplification at 14q12 may be characteristic for HCCs. No significant difference in chromosomal aberrations was noted between carcinomas associated with HCV-infection in our study and those reported earlier in HCCs infected with hepatitis B virus (HBV), indicating that both HBV- and HCV-related carcinomas may progress through a similar cascade of molecular events.     

Intracellular site of "Ca reversal": inhibition of uterine smooth muscle contraction in Ca-free medium by a minute amount of Ca ion released from mitochondria by drugs

Sodium azide (NaN3) and dinitrophenol (DNP) at low concentrations caused "Ca reversal", i.e. inhibition of oxytocin-induced tonic contraction of estrogen-dominated rat uterine smooth muscle in Ca-free solution. This inhibition was not due to depletion of ATP by NaN3 or DNP. Higher concentrations of NaN3 and DNP caused additional contraction. NaN3 and DNP dose-dependently released Ca ion from mitochondria isolated from the estrogen-dominated rat uterine smooth muscle in vitro. Electron microscopic studies have shown that in estrogen-dominated rat uterine smooth muscle cells, cytoplasmic membranes proliferate, resulting in compartmentalization of the myofilament-sarcoplasmic system and its separation from the receptor-effector system in the surface folds of the plasma membranes, which also contain some mitochondria. It is proposed that low concentrations of NaN3 and DNP release a small amount of Ca ion from these outer mitochondria and this Ca ion acts on the intracellular "site of Ca reversal" to induce reversal, i.e. inhibition. Higher concentrations of NaN3 and DNP are proposed to release a large amount of Ca ion from the central mitochondria near myofilaments and so induce contraction. The "site of Ca reversal" was shown to be intracellular as our previous postulation.     

Acute effects of statin on reduction of angiopoietin-like 2 and glyceraldehyde-derived advanced glycation end-products levels in patients with acute myocardial infarction: a message from SAMIT (Statin for Acute Myocardial Infarction Trial)

Experimental ischemia–reperfusion models have shown that 3-hydroxy-3methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, statins, have cardioprotective effects. SAMIT (Statin Acute Myocardial Infarction Trial) is a multicenter prospective open randomized trial, designed to evaluate the effects of statin treatment from the earliest stage on cardioprotection in patients with acute myocardial infarction (AMI). Patients were randomly assigned to receive atorvastatin (initial dose of 40 mg at admission followed by the maintenance dose of 10 mg/day for 30 days) or not (control), and then immediately underwent percutaneous coronary intervention (PCI) for the culprit lesion. The primary endpoints were infarct size and left ventricular function. The secondary endpoints were major adverse cardiac and cerebrovascular events (MACCE) and various biomarkers. There were no significant differences in baseline characteristics between 2 groups of the statin treatment group and the control group. The left ventricular ejection fraction increased at 6 months after the onset of AMI, compared with the baseline level in the atorvastatin group (P < 0.05), while it did not change in the control group. Although there were no significant differences in the MACCE, the changes in the levels of angiopoietin-like protein 2 (ANGPTL2) (P < 0.05), and glyceraldehyde-derived advanced glycation end-products, (TAGE) (P < 0.01) were suppressed at 2 weeks in the atorvastatin group, compared with the control group. Statin therapy started early after the onset reduced the levels of ANGPTL2 and TAGE, and thus, might have cardioprotective effects in patients with AMI.     

Rapid ecotoxicological bioassay using delayed fluorescence in the marine cyanobacterium <Emphasis Type="Italic">Cyanobium</Emphasis> sp. (NIES-981)

The use of delayed fluorescence intensity as an endpoint for rapid estimation of the effective concentration (EC_x) has been reported as an alternative to standard growth inhibition (at 72?h after exposure) in some algal species including Pseudokirchneriella subcapitata. In marine algae, although an approach of bioassaying using delayed fluorescence measurements has not been performed yet, its development would provide many benefits for marine environmental risk assessment. In this study, we selected marine cyanobacterium Cyanobium sp. (NIES-981) as our test algal species and demonstrated that this species is valid for the standard growth inhibition test based on criteria provide by Organization for Economic Co-operation and Development guidelines. Furthermore, standard inhibition tests and shorter period test using DF were performed in NIES-981 using five chemicals (3,5-DCP, simazine, diflufenican, K₂Cr₂O₇, and CuSO₄), and their EC₅₀ and low-toxic-effect values (EC₁₀, EC₅, and NOEC) were determined from two dose-response curves. Based on comparisons of the two dose-response curves and the EC₅₀ values, we conclude that DF intensity is useful as an endpoint for rapid estimation of EC₅₀ in NIES-981.