Molecular mechanism of DNA replication-coupled inactivation of the initiator protein in Escherichia coli: interaction of DnaA with the sliding clamp-loaded DNA and the sliding clamp-Hda complex

In Escherichia coli, the ATP-DnaA protein initiates chromosomal replication. After the DNA polymerase III holoenzyme is loaded on to DNA, DnaA-bound ATP is hydrolysed in a manner depending on Hda protein and the DNA-loaded form of the DNA polymerase III sliding clamp subunit, which yields ADP-DnaA, an inactivated form for initiation. This regulatory DnaA-inactivation represses extra initiation events. In this study, in vitro replication intermediates and structured DNA mimicking replicational intermediates were first used to identify structural prerequisites in the process of DnaA-ATP hydrolysis. Unlike duplex DNA loaded with sliding clamps, primer RNA-DNA heteroduplexes loaded with clamps were not associated with DnaA-ATP hydrolysis, and duplex DNA provided in trans did not rescue this defect. At least 40-bp duplex DNA is competent for the DnaA-ATP hydrolysis when a single clamp was loaded. The DnaA-ATP hydrolysis was inhibited when ATP-DnaA was tightly bound to a DnaA box-bearing oligonucleotide. These results imply that the DnaA-ATP hydrolysis involves the direct interaction of ATP-DnaA with duplex DNA flanking the sliding clamp. Furthermore, Hda protein formed a stable complex with the sliding clamp. Based on these, we suggest a mechanical basis in the DnaA-inactivation that ATP-DnaA interacts with the Hda-clamp complex with the aid of DNA binding.     

Potential oscillation elicited by i.v. olfaction and its applicability as an objective clinical olfaction test

OBJECTIVE: Alinamin has long been applied in Japan for testing i.v. olfaction and to diagnose olfactory disorders. The test is subjective, each subject being asked about the presence or absence of olfaction. The credibility of the answers is highly questionable in some cases; as a result, the reliability of the test is poor. Recent studies demonstrated an induced electric potential in the scalp during i.v. olfactory testing. Some patients complain of the pain of the injection during i.v. olfactory testing; therefore, the effect of this pain must be considered with respect to measurement of the i.v. olfaction-elicited potential (IVOP). MATERIAL AND METHODS: This investigation involved 179 subjects with various olfaction levels. Each subject received an Alinamin injection; the elicited potential amplitude was compared before and after the injection and the increasing ratio (IR) was computed. Gender, age, level of olfactory disorder, the presence or absence of olfaction and the presence or absence of the pain of injection were considered as factors affecting IR. RESULTS: IR showed significant increases in groups characterized by the presence of olfaction as well as in groups reporting pain of injection. The test subjects were further divided into four groups based on their olfaction and pain of injection patterns as follows: Group A, no smell and no pain; Group B, smell and no pain; Group C, no smell and pain; and Group D, smell and pain. Subjects exhibiting no recognizable olfaction or pain of injection (Group A) revealed no increase in IVOP following injection. Subjects with either recognizable olfaction or pain of injection (Groups B and C) exhibited a slight increase in IVOP following injection. Subjects with both noticeable olfaction and pain of injection (Group D) demonstrated a significant increase in IVOP following the injection with a very high value of IR (>2). Furthermore, there were significant differences between the four groups in terms of IR level, with the exception of Groups B and C. CONCLUSIONS: Olfaction is largely involved with the generation of IVOP. However, pain resulting from injection of Alinamin is considered to be a significant factor. IVOP showed significant effectiveness for diagnosing olfactory disorders in cases who did not experience pain of injection.     

Inactivating and non-inactivating potassium currents in isolated inner hair cells from guinea pig cochlea

Using conventional whole-cell voltage-clamp recordings, we examined the 4-aminopyridine (4-AP)- and tetraethylammonium (TEA)-sensitive K(+) currents in the cochlear inner hair cells (IHCs) of guinea pigs. 4-AP-sensitive currents were activated slowly and sustained the same current level, whereas TEA-sensitive currents were activated rapidly, followed by inactivation. The inactivation time course of TEA-sensitive currents was voltage-dependent, becoming faster at more depolarized levels. The inactivation of TEA-sensitive currents almost recovered within 5 ms. 4-AP- and TEA-sensitive K(+) currents coexisted in the same IHC.     

Oral ADSORBENT AST-120 decreases carotid intima-media thickness and arterial stiffness in patients with chronic renal failure

BACKGROUND/AIM: Intima media thickness (IMT) and stiffness of the carotid arteries is related to coronary artery disease, and chronic renal failure patients are at high risk for such diseases. An oral adsorbent, AST-120 (Kremezin; Kureha Chemical Industry, Tokyo, Japan), can delay the progression of chronic renal failure in undialyzed uremic patients. The aim of the present study was to determine whether AST-120 affects carotid artery IMT and pulse wave velocity (PWV) in patients with chronic renal failure not undergoing dialysis. METHODS: Fifty patients with non-diabetic chronic renal failure were randomly divided into two groups: 30 patients (18 men and 12 women; mean age 53.5 years; mean serum creatinine 3.2 mg/dl) who were given AST-120 (6.0 g/day) and 20 patients (12 men and 8 women; mean age 52.0 years; mean serum creatinine 3.5 mg/dl) who were not given AST-120. Thirty healthy age-matched subjects (18 men and 12 women; mean age 51.5 years; mean serum creatinine 0.9 mg/dl) were also included. The treatment period was 24 months. IMT and arterial stiffness were measured before and after treatment. RESULTS: The slope of the reciprocal serum creatinine concentration over time became significantly less steep in the AST-120 group than in the non-AST-120 group (p < 0.001). Before treatment, carotid artery IMT differed little between the AST-120 group (0.90 +/- 0.22 mm) and the non-AST-120 group (0.88 +/- 0.20 mm). IMT in these two groups was significantly greater than IMT in the control group (0.64 +/- 0.14 mm) (p < 0.01). Carotid IMT in the AST-120 group decreased slightly but not significantly to 0.84 +/- 0.20 mm after 12 months and then significantly after 24 months to 0.78 +/- 0.18 mm (p < 0.05). Carotid IMT in the non-AST group showed little change throughout the experimental period. PWV differed little between the AST-120 group (1,980 +/- 330 cm/s) and the non-AST group (1,940 +/- 360 cm/s) before treatment. PWV values in these two groups were significantly greater than PWV in the control group (1,280 +/- 240 cm/s) (p < 0.01). After 12 and 24 months, PWV in the AST-120 group decreased significantly to 1,840 +/- 280 cm/s (p < 0.05) and to 1,780 +/- 260 cm/s (p < 0.05), respectively; however, PWV in the non-AST group showed a slight increase during the experimental period. CONCLUSION: The data suggest that AST-120 may reduce arterial stiffness and IMT in non-diabetic chronic renal failure patients before dialysis.     

A telemedicine system for collaborative work on radiographic coronary video-images

We designed a telemedicine system that supports collaborative work on video-images from coronary cine-angiograms and cine-ventriculograms. Two workstations were connected by ISDN at 128 kbit/s. Coronary cine-angiogram and cine-ventriculogram video-images were transmitted to an image examination workstation before the images were required for analysis (i.e. in a 'pre-fetch' operation). If one workstation changed even a single frame in a video-sequence, then the change was transmitted to the other workstation, rather than the entire video-sequence. To evaluate the performance of the system, the diagnoses made by five cardiology specialists were compared with those recorded in the original report. Coronary cine-angiogram and cine-ventriculogram video-images from 27 patients were used. The cardiologists, who were blinded to the patient information, regenerated the video-images on the workstation and assessed coronary arterial stenosis and wall motion. The kappa scores for clinician agreement with the original reports for ratings of stenosis were 0.54-0.66 and for wall motion were 0.41-0.69. The system performed reasonably reliably and it therefore appears likely to be useful in telemedicine work.     

Impact of health education and screening over all-cause mortality in Japan: evidence from a cohort study during 1984-2002

BACKGROUND: Health education and screening are two components of preventive health services in Japan since 1983. This study investigated their relationships with all-cause mortality as they are studied insufficiently in Hokkaido, Japan. METHODS: This study enrolled 1,532 men and 1,653 women aged 40-97 years from 1,702 randomly selected households of 60 areas during 1984-1985 and followed them until 2002. At baseline survey, staffs of 45 health centers collected sociodemographic, medical, behavioral, and dietary information including health education and screening from study subjects with informed consent. RESULTS: For men, age-adjusted Cox proportional hazard model indicated lower mortality for those who received health education (RR = 0.76, P < 0.01) and screening (RR = 0.83, P < 0.05) than those who did not. Health education showed lower mortality even after adjusting for many variables. Similarly for women, health education (RR = 0.66, P < 0.01) and screening (RR = 0.64, P < 0.001) revealed lower age-adjusted mortality. Almost similar results were found for both services when models were adjusted for many variables and when the deaths including lost to follow-up cases of the first 4 years of baseline survey are excluded. CONCLUSIONS: This study shows protective effects of health education and screening over all-cause mortality for both sexes. However, further studies are needed to confirm the results.     

Associations between dietary intake and urinary excretion of sodium, potassium, phosphorus, magnesium, and calcium

OBJECTIVES AND METHODS: The associations between dietary intake and urinary excretion of sodium (Na), potassium (K), calcium (Ca), magnesium (Mg), and phosphorus (P), and the major dietary sources derived from the urinary minerals were studied in a nutritional survey of 219 Japanese females aged 27-84 years, who completed anthropometric measurements, a one-day dietary record, and a 24 hr urine collection. RESULTS: The minerals excreted in the urine were significantly and positively correlated with each other, in which Na excretion was correlated with K and Ca excretion (r = 0.490 and r = 0.482, respectively, p < 0.01) and Ca excretion was correlated with Mg excretion (r = 0.526, p < 0.01). The ratios of urinary exertion to dietary intake of Na, K, Ca, Mg, and P were 81.5%, 62.7%, 24.5%, 21.7%, and 56.1%, respectively. The dietary intake and the urinary excretion of the minerals expressed per body weight (kg) were significantly and positively correlated (Na, r = 0.267; K, r = 0.460; Ca, r = 0.181; Mg, r = 0.245; P, r = 0.351, p < 0.01). Further examinations using chief component analysis for food intake showed several significant positive correlations, including between Na intake and the intake of vegetables, noodles, and seasonings (r = 0.332-0.381, p < 0.01); between K, Mg and P intake and the intake of vegetables, fruits, and potatoes (r = 0.332-0.533, p < 0.01); and between Ca intake and the intake of bread and dairy foods (r = 0.428, p < 0.01). In addition, significant positive associations were found between Na excretion and the intake of confectionaries, nuts, and seeds (r = 0.223, p < 0.01). Weak correlations were also found between K excretion and the intake of vegetables (r = 0.296, p < 0.01); between Ca and P excretion and the intake of meat, oil, and fats (r = 0.135, P < 0.05; r = 0.193, P < 0.01, respectively), and between Mg excretion and the intake of bread and dairy foods (r = 0.137, P < 0.05). CONCLUSIONS: Findings from this study indicate that, while urinary excretion of Ca and Mg is unlikely to be a reliable biochemical marker of dietary intake, the levels of urinary excretion of Na, K, and P can be reflective of the intake of salt, vegetables, and meats, respectively. The urinary excretion of the minerals, particularly Na, K, and Ca, may be highly linked to salt intake in Japanese females.     

Effects of SEA0400, a novel inhibitor of the Na+/Ca2+ exchanger, on myocardial stunning in anesthetized dogs

Activation of the Na+/Ca2+ exchanger may contribute to Ca2+ overload during reperfusion after transient ischemia. We examined the effects of 2-[4-[(2,5-difluorophenyl) methoxy]phenoxy]-5-ethoxyaniline (SEA0400), a selective inhibitor of Na+/Ca2+ exchange, on a canine model of ischemia/reperfusion injury (myocardial stunning). Myocardial stunning was induced by a 15-min occlusion of the left anterior descending coronary artery followed by a 4-h reperfusion in anesthetized open-chest dogs. Reperfusion gradually restored myocardial percent segment shortening but remained depressed during a 4-h reperfusion period. A bolus intravenous injection of SEA0400 (0.3 or 1.0 mg/kg), given 1 min before reperfusion, improved significantly the recovery of percent segment shortening in the ischemic/reperfused myocardium. SEA0400 did not affect the hemodynamics and electrocardiogram parameters. In addition, SEA0400 did not affect reperfusion-induced change in coronary blood flow. These results suggest that the Na+/Ca2+ exchanger is involved in the stunned myocardium of dogs after reperfusion, and that SEA0400 has a protective effect against myocardial stunning in dogs.