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The etiology of peripartum cardiomyopathy (PPCM) is unknown. Therefore, we evaluated the etiology of patients clinically diagnosed with PPCM using endomyocardial biopsy. We studied five patients diagnosed with PPCM following endomyocardial biopsy (age, 28–42 years; mean age, 35 years). Biopsied samples were evaluated using microscopy, including immunostaining and electron microscopy. The pathological findings were as follows: myocardial hypertrophy, myocardial fibrosis, and cell infiltration. Two patients were diagnosed with lymphocytic myocarditis, one with eosinophilic myocarditis, one with hypertensive heart disease, and one with a combination of hypertension and myocarditis. Endomyocardial biopsy suggested that the causes of PPCM were varied and related to myocarditis and myocardial overload due to hypertension. [ABSTRACT FROM AUTHOR]

Restrictive cardiomyopathy (RCM) is a rare primary myocardial disease, and its pathological features are yet to be determined. Restrictive cardiomyopathy with MHY7 mutation was diagnosed in a 65-year-old Japanese woman. Electron microscopy of a myocardial biopsy revealed electron-dense materials resulting from focal myocyte degeneration and necrosis as well as tubular structures and pseudo-inclusion bodies in some nuclei. These features may be associated with the pathogenesis of RCM. [ABSTRACT FROM AUTHOR]

A 16-year-old Japanese man was admitted to our hospital because of syncope during exercise. His father and his younger brother had permanent pacemaker implantation because of sick sinus syndrome. Several examinations revealed first-degree atrioventricular block, complete right bundle branch block, sick sinus syndrome, and ventricular tachycardia with normal cardiac function. As no abnormalities were evident on coronary angiography, right ventricular endomyocardial biopsy was performed. It showed myocardial disarrangement and lipofuscin accumulation in hypertrophic myocytes. Moreover, electron microscopy showed a few degenerative myocytes, Z-band streaming, disarrangement, increased small capillaries with Weibel-Palade bodies in endothelial cells, and endothelial proliferations. Genetic analysis of the proband, his father, and his younger brother revealed a missense mutation, D1275N, in SCN5A, a gene which encodes sodium ion channel protein, are related to cardiomyopathy and arrhythmia. The proband was diagnosed with a cardiac conduction defect (CCD) and underwent permanent pacemaker implantation. These pathological findings suggest various myocardial changes presented in CCD patients with a missense mutation, D1275N, in SCN5A. [ABSTRACT FROM AUTHOR]

Chronic myocarditis is sometimes difficult to diagnose using several clinical diagnostic modalities. A 43‐year‐old Japanese man was admitted to our hospital with heart failure due to a diffusely hypokinetic left ventricle. No abnormal accumulation was seen on 18F‐fluorodeoxyglucose positron emission tomography/computed tomography. Coronary angiography showed no abnormalities. Endomyocardial biopsy was performed on suspicion of dilated cardiomyopathy, revealing diffuse cell infiltration (more T lymphocytes associated with macrophages than B cells on immunohistochemical staining), myocyte damage, and replacement fibrosis. The pathological diagnosis of biopsy specimen was difficult to differentiate between chronic myocarditis and inflammatory dilated cardiomyopathy without immunohistochemistry. Endomyocardial biopsy offers one of the most useful methods for diagnosing chronic myocarditis. [ABSTRACT FROM AUTHOR]

Vibrio vulnificus , the causative agent of serious, often fatal, infections in humans, requires iron for its pathogenesis. As such, it obtains iron via both vulnibactin and heme-mediated iron-uptake systems. In this study, we identified the heme acquisition system in V . vulnificus M2799. The nucleotide sequences of the genes encoding heme receptors HupA and HvtA and the ATP-binding cassette (ABC) transport system proteins HupB, HupC, and HupD were determined, and then used in the construction of deletion mutants developed from a Δ ics strain, which could not synthesize vulnibactin. Growth experiments using these mutants indicated that HupA and HvtA are major and minor heme receptors, respectively. The expressions of two proteins were analyzed by the quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). Furthermore, complementation analyses confirmed that the HupBCD proteins are the only ABC transport system shared by both the HupA and HvtA receptors. This is the first genetic evidence that the HupBCD proteins are essential for heme acquisition by V . vulnificus . Further investigation showed that hupA , hvtA , and hupBCD are regulated by Fur. The qRT-PCR analysis of the heme receptor genes revealed that HupR, a LysR-family positive transcriptional activator, upregulates the expression of hupA , but not hvtA . In addition, ptrB was co-transcribed with hvtA , and PtrB had no influence on growth in low-iron CM9 medium supplemented with hemin, hemoglobin, or cytochrome C. [ABSTRACT FROM AUTHOR]

The article presents a case study of a 85 year old woman who underwent a nephrostomy for right hydronephrosis. It notes that after the surgery she suffered acute heart failure and several tests revealed that she was diagnosed with midventricular Takotsubo (or stress-induced) cardiomyopathy (TC). The article discusses that the patient's condition improved due to the infusion of landiolol and was released from the hospital after two days.

Protein introduction into cells is more difficult in plants than in mammalian cells, although it was reported that protein introduction was successful in shoot apical meristem and leaves only together with a cell-penetrating peptide. In this study, we tried to introduce superfolder green fluorescent protein (sGFP)-fused to adenylate cyclase as a reporter protein without a cell-penetrating peptide into the cells of tobacco leaves by treatment with atmospheric non-thermal plasmas. For this purpose, CO2 or N2 plasma was generated using a multi-gas plasma jet. Confocal microscopy indicated that sGFP signals were observed inside of leaf cells after treatment with CO2 or N2 plasma without substantial damage. In addition, the amount of cyclic adenosine monophosphate (cAMP) formed by the catalytic enzyme adenylate cyclase, which requires cellular calmodulin for its activity, was significantly increased in leaves treated with CO2 or N2 plasma, also indicating the introduction of sGFP-fused adenylate cyclase into the cells. These results suggested that treatment with CO2 or N2 plasma could be a useful technique for protein introduction into plant tissues. [ABSTRACT FROM AUTHOR]

To identify mechanisms underlying the bacterial inactivation process by atmospheric nonthermal plasma using a unique plasma jet that can generate various gas plasmas, Staphylococcus aureus were irradiated with carbon dioxide plasma, which produces a large amount of singlet oxygens, and nitrogen plasma, which produces a large amount of OH radicals. And damaged areas of plasma-treated bacteria were observed by field emission scanning electron microscopy, transmission electron microscopy, and atomic force microscopy. As a result, bacteria were damaged by both gas plasmas, but the site of damage differed according to gas species. Therefore, it suggests that singlet oxygen generated by carbon dioxide plasma or other reactive species caused by singlet oxygen contributes to the damage of internal structures of bacteria through the cell wall and membrane, and OH radicals generated by nitrogen plasma or other reactive species derived from OH radicals contribute to damage of the cell wall and membrane. [ABSTRACT FROM AUTHOR]

A 13-year-old female with Noonan syndrome had been diagnosed with hypertrophic cardiomyopathy, and she died of heart failure at the age of 25 years. Light microscopic and electron microscopic examination of her biopsied myocardium and autopsy heart showed myocardial fragmentation associated with Z-band disruption as well as myocardial hypertrophy and disarray with interstitial fibrosis. Myocardial fragmentation associated with Z-band disruption may be related to the progression of cardiac dysfunction. [ABSTRACT FROM AUTHOR]

An atmospheric pressure plasma source in which the gas temperature can be accurately controlled from below freezing point up to a high temperature has been developed. In general plasma devices, an electrical discharge is passed through a gas at about room temperature to generate plasma, so the plasma is at a temperature higher than room temperature; moreover, the gas temperature is determined by the discharge condition such as discharge power and plasma gas flow rate, so accurate temperature control is difficult. In the plasma source that has been developed in this research, the gas that is to be supplied to the discharge unit is first cooled using a gas cooler and then heated by a heater. The gas temperature of the produced plasma is measured, and feedback is sent to the heater. Thus, plasma at a desired temperature can be generated. Gas temperature control of the plasma over a range from −54 °C to +160 °C with a standard deviation of 1 °C was realized. Spectroscopic characteristics of generated plasma were investigated. This plasma source/technique will realize that effective plasma is applicable for heat-sensitive materials such as paper, textile, polymer, and especially human tissue. Furthermore, it enables us to generate the plasma at optimal gas temperature for chemical reaction of each plasma treatment. [ABSTRACT FROM PUBLISHER]

Vibrio vulnificus , an opportunistic marine bacterium that causes a serious, often fatal, infection in humans, requires iron for its pathogenesis. This bacterium exports vulnibactin for iron acquisition from the environment. The mechanisms of vulnibactin biosynthesis and ferric-vulnibactin uptake systems have recently been reported, while the vulnibactin export system has not been reported. Mutant growth under low-iron concentration conditions and a bioassay of the culture supernatant indicate that the VV1_0612 protein plays a crucial role in the vulnibactin secretion as a component of the resistance-nodulation-division (RND)-type efflux system in V . vulnificus M2799. To identify which RND protein(s) together with VV1_0612 TolC constituted the RND efflux system for vulnibactin secretion, deletion mutants of 11 RND protein-encoding genes were constructed. The growth inhibition of a multiple mutant (Δ11) of the RND protein-encoding genes was observed 6 h after the beginning of the culture. Furthermore, Δ VV1_1681 exhibited a growth curve that was similar to that of Δ11, while the multiple mutant except Δ VV1_1681 showed the same growth as the wild-type strain. These results indicate that the VV1_1681 protein is involved in the vulnibactin export system of V . vulnificus M2799. This is the first genetic evidence that vulnibactin is secreted through the RND-type efflux systems in V . vulnificus . [ABSTRACT FROM AUTHOR]

Background: We investigated the clinical course of complete right bundle branch block (RBBB) or RBBB with axis deviation (AD) in terms of subsequent pacemaker implantation for high-degree atrioventricular (AV) block or sick sinus syndrome (SSS). Methods and results: Among the 16,170 atomic-bomb survivors in our biennial health examination between July 1967 and December 2010, we detected 520 newly-acquired RBBB subjects with no organic heart disease, and selected 1038 age- (at RBBB diagnosis) and sex-matched subjects without RBBB to serve as comparison subjects. Multivariate Cox regression analysis was used to estimate the hazard ratios (HRs) for the risk of pacemaker implantation due to all causes, AV block or SSS between RBBB and comparison subjects and between RBBB subjects with and without AD. The risk of pacemaker implantation for RBBB was 4.79 (95% confidence interval [CI] 1.89–12.58; P =0.001), 3.77 (95% CI, 1.09–13.07; P =0.036), and 6.28 (95% CI, 1.24–31.73, P =0.026) when implantation was for all causes, AV block and SSS, respectively. RBBB subjects with AD had a higher risk for all-cause pacemaker implantation than subjects without AD (HR, 3.03; 95% CI, 1.00–9.13, P =0.049). RBBB subjects with AD were younger than subjects without AD at the time of RBBB diagnosis (59.4±7.6 vs 74.4±3.1years old, P =0.019), and their progression from diagnosis to pacemaker implantation took longer (15.1±6.6 vs 6.4±3.0years, P =0.032). Conclusions: RBBB, especially with AD, progresses to AV block and SSS that requires pacemaker implantation; the mechanisms by which the conduction defect progresses differ among patients with and without AD. [ABSTRACT FROM AUTHOR]

Aim We evaluated the effect of pimobendan, a positive inotropic agent, in elderly patients with frequent readmission as a result of heart failure despite conventional therapy. Methods Pimobendan was given to five male patients with severe chronic heart failure ( New York Heart Association class III- IV) (age range 69-89 years; mean 78 ± 8 years; ischemic cardiomyopathy in three cases, dilated cardiomyopathy in two cases) who required repeated admission for heart failure despite conventional therapy with angiotensin inhibitors, beta-blockers, diuretics and anti-arrhythmic agents. After the addition of pimobendan at a dose of 1.25-3.75 mg/day, we evaluated serum levels of brain natriuretic peptide ( BNP), left ventricular ejection fraction ( LVEF), septal e′ and left ventricular end-diastolic diameter ( LVDD) by echocardiography, as well as readmission rates for more than 2 years. Results The serum level of BNP significantly decreased after treatment with pimobendan, although its level returned to pretreatment levels after 2 years. LVEF significantly improved after the treatment, with the improvement continuing beyond the 2 years, although LVDD did not change after treatment. Septal e′ significantly improved after the treatment, although its level returned to pretreatment levels at 2 years after the treatment. Readmission rates significantly decreased for 2 years after the treatment, although one patient required cardiac resynchronization therapy for severe heart failure, and another patient required cardiac pacemaker implantation for sick sinus syndrome 2 years after adding pimobendan. Conclusions Pimobendan in conjunction with conventional therapy for heart failure decreases the readmission rate in elderly patients with severe heart failure for at least 2 years. Geriatr Gerontol Int 2014; 14: 109-114. [ABSTRACT FROM AUTHOR]

Abstract: Vibrio vulnificus, an opportunistic marine bacterium that causes a serious, often fatal, infection in humans, requires iron for its pathogenesis. This bacterium uses iron from the environment via the vulnibactin-mediated-iron-uptake system. In this study, we constructed the deletion mutants of the genes encoding the proteins involved in the vulnibactin-mediated-iron-uptake system, isochorismate synthase (ICS), vulnibactin utilization protein (VuuB), periplasmic ferric-vulnibactin binding protein (FatB), and ferric-vulnibactin receptor protein (VuuA). The Δics and ΔvuuA mutants were unable to grow under low-iron concentration conditions compared with the isogenic wild-type, indicating that the involvement of ICS in the vulnibactin biosynthesis pathway and uptake of ferric-vulnibactin through the VuuA receptor protein are essential for V. vulnificus M2799 growth under low-iron concentration conditions. Similar growth impairment was also observed in ΔfatB, with growth recovery of this mutant observed 6 h after the beginning of the culture. These results indicate that there must be other periplasmic ferric-vulnibactin binding proteins in V. vulnificus M2799 that complement the defective fatB gene. Complementary growth studies confirmed that VatD protein, which functions as a periplasmic ferric-aerobactin binding protein, was found to participate in the ferric-vulnibactin uptake system in the absence of FatB. Furthermore, the expression of ics, vuuB, fatB, vuuA, and vatD genes was found to be regulated by iron and the ferric uptake regulator. [Copyright &y& Elsevier]

Microbes with smaller genomes would be better chassis for analysis, design, and improvement in the fields of metabolic engineering, synthetic biology, and molecular breeding. To create an Escherichia coli strain with a smaller genome, we used a stepwise genome reduction approach. Beginning with strain MGF-01, which has a genome of 3.62 megabase pairs (Mbp), we generated two E. coli K-12 strains without any insertion sequence (IS), DGF-327 and DGF-298, with reduced genome sizes of 3.27 and 2.98 Mbp, respectively. During the strain construction, intrinsic mutations of ilvG and rph were functionally restored to accelerate initial growth after inoculation. The genomes of the two strains were sequenced, and their structures were confirmed. Both strains showed no auxotrophy, and had better growth fitness, especially in the initial phase, and better cell yield in a rich medium than the wild type K-12 strain. Transcriptome analysis revealed that ibpAB and lon, which encode a heat-shock chaperone and a protease for abnormal proteins, respectively, are down-regulated in DGF strains, compared to the ancestral strains with larger genomes. We concluded that down-regulation of the genes encoding chaperones and proteases is one of the factors that improve the fitness of DGF strains. The DGF strains with fewer genes and better cell yield will be good hosts for applications. [ABSTRACT FROM AUTHOR]

Nucleocytoplasmic transport of proteins across the nuclear pore complex (NPC), mediated by the nuclear localization signal (NLS) and the nuclear export signal (NES), is a vital homeostatic process in eukaryotic cells and also in mitogen-activated protein kinase (MEK) signaling molecule in tumor cell proliferation. Some viruses, including the influenza virus and HIV-1, also employ this nuclear export mechanism during their life cycle. Hence, drugs that control nucleocytoplasmic transport of proteins are putative candidate antivirals or anti-cancer agents. Thus, we previously developed a GFP/NES-MDCK reporter cell system for screening novel nuclear export inhibitors. NES signal-conjugated GFP accumulates in the nucleus in the presence of the nuclear export inhibitor leptomycin B (LMB). In this study, a stable GFP/NLS/NES fusion protein-expressing cell line was established, and its potential as a reporter was evaluated. The GFP/NLS/NES-MDCK cell line demonstrates improved nuclear accumulation of GFP in a time-course treatment with LMB. In addition, the dose-response data demonstrated superior sensitivity of GFP/NLS/NES-MDCK over GFP/NES-MDCK cells. As low as 0.01 ng/mL LMB is sufficient to cause accumulation of the GFP fusion protein in the nucleus in GFP/NLS/NES-MDCK cells, while at least 1 ng/mL of LMB is needed for the accumulation of GFP fusion protein in the nucleus of GFP/NES-MDCK cells. These results indicate that the newly established GFP/ NLS/NES-MDCK cell line is a potentially powerful tool to screen for novel nuclear export inhibitors. [ABSTRACT FROM AUTHOR]

Various oxidative stress markers have been measured to evaluate the status of heart failure (HF). However, the relationships between these markers and the aetiology of HF have not been fully investigated. This study compared 8-hydroxy-2′-deoxyguanosine (8-OHdG) and biopyrrins levels in patients with ischemic and non-ischemic HF. Study subjects were divided into a coronary artery disease (CAD) group ( n=70), a non-CAD group ( n=61) and a control group ( n=33). In the CAD group, 8-OHdG and biopyrrins levels increased with the severity of the New York Heart Association (NYHA) functional class and log BNP levels correlated with 8-OHdG and biopyrrins levels. However, non-CAD patients with NYHA class III/IV had significantly lower 8-OHdG levels than CAD patients with NYHA class III/IV and the levels did not correlate with log BNP levels. In the CAD group, 8-OHdG levels reflected the severity of atherosclerosis. These results indicate that the properties of oxidative stress markers should be carefully taken into consideration for the assessment of HF status. [ABSTRACT FROM AUTHOR]

The article presents a study which designs and constructs six major toxin-antitoxin (TA) disruptants of Escherichia coli K-12 W3110. Among the cultivated disruptants, Δ high-frequency-persistence mutants BA (ΔhipBA) showed a longer life span than other disruptants. The said disruptant may also function as a bacteriostatic system based on the physiological analysis.

Abstract: Endothelial dysfunction precedes the development of clinical atherosclerosis. A decrease in endothelium-derived nitric oxide activity shows impaired vasodilator function and causes arterial intimal hyperplasia or thickening. Others and we have reported the close relation of endothelial function in brachial artery and coronary artery. To compare the flow-mediated endothelium-dependent vasodilation (FMD) in brachial artery and the intima+media area in coronary artery, we measured brachial artery vasodilator response following transient occlusion with high resolution ultrasound transducer and intima+media area in coronary arteries with intravascular ultrasound in 25 patients with normal coronary angiograms (age 61.6±8.7 years old, men 14 and women 11). FMD was measured at least 7 days after the cessation of all vasodilators. The mean FMD of 25 patients was 3.83±2.38%, the mean intima+media area in coronary arteries of 25 patients was 39.9±15.5% of total vessel wall. FMD has a close negative relation with the largest percent intima+media area (r =−0.77, p <0.01). Especially, the patients whose FMD was less than 3.83% had larger percent intima+media area than those whose FMD was 3.83% or more (48.7±10.7% versus 30.3±14.2%, p <0.01). There is an intima+media thickening even in the patients who had normal coronary angiograms, and that the percent intima+media area correlated with FMD. The measurement of FMD is useful for screening the coronary artery intima+media thickening noninvasively. The pathogenesis of acute coronary syndrome has been reported to be the plaque rupture even in the patients with normal coronary angiograms. Thus, we must pay much attention in the patients with impaired FMD even in the normal coronary angiograms. [Copyright &y& Elsevier]

Abstract: The role of one of the major outer membrane proteins, rOmpB, of spotted fever group rickettsiae was examined. Antibodies generated against native rOmpB inhibited plaque formation by Rickettsia japonica in Vero cells when applied at the time of inoculation of the rickettsiae. However, antibodies to heat-denatured rOmpB did not. Moreover, the soluble recombinant rOmpB also inhibited plaque formation to some extent. Thus it seems that rOmpB functions at least in the adherence of rickettsiae to host cells. To obtain direct evidence of its function in the adherence to and invasion of Vero cells, we generated Escherichia coli transformed by the vector pET-22b(+) inserted with the ompB open reading frame of R. japonica. The recombinant bacteria expressed a 165-kDa protein consistent with the precursor of rOmpB. The protein reacted with monoclonal antibodies to heat-labile epitopes of rOmpB. Immunofluorescence of the recombinant bacteria demonstrated surface expression of the protein. It was shown by light microscopy and transmission and scanning electron microscopy that the bacteria adhered to and invaded Vero cells. Thus, although the recombinant precursor rOmpB was not processed on the outer membrane of E. coli, it functions during these steps. The manner of entry was similar to that of rickettsiae although at a slower rate. [Copyright &y& Elsevier]

The genes of cytochrome bd-encoding cydAB were identified from a deep-sea bacterium Shewanella violacea DSS12. These showed significant homologies with known cydAB gene sequences from various organisms. Additionally, highly conserved regions that are important for the enzymatic function were also conserved in cydA of S. violacea. Based on the results, transcriptional analysis of cydAB operon and cydDC operon (required for assembly of cytochrome bd) of S. violacea in microaerobic condition was performed under the growth condition of various pressures. The gene of cydA was expressed even under the condition of atmospheric pressure and its expression was enhanced with pressurization. On the other hand, the expression of cydC was strongly depressed under the condition of atmospheric pressure compared with the case under high pressure. It appeared spectrophotometrically that loss of cytochrome bd in S. violacea under atmospheric pressure shown in previous study is caused mainly by the loss of cydDC. Further, under the growth condition of atmospheric pressure, either less amount or no d-type cytochrome was expressed compared with the case of high-pressure condition even if the organism was grown under alkaline condition or in the presence of uncoupler, which are the inducible condition of d-type cytochrome in Escherichia coli. These results suggested that the significant amount of d-type cytochrome expression is specific event under the growth condition of high pressure. [ABSTRACT FROM AUTHOR]

Plasma levels of human thioredoxin are indicative of the responses against oxidative stress. We measured the plasma thioredoxin levels in patients with unstable angina in order to examine the relationships between subsequent clinical course and plasma thioredoxin levels before and after treatment for unstable angina. Blood was sampled both on admission and after treatment in 44 patients with unstable angina. In addition, blood samples were obtained from 41 patients with stable exertional angina and 41 patients with chest pain syndrome after admission. The plasma levels of thioredoxin were the highest in the unstable angina group among three groups (p<0.001). Treatment of unstable angina decreased the plasma thioredoxin levels (p<0.01). We divided the patients with unstable angina into two groups according to the plasma thioredoxin levels on admission and after treatment. There was a significant difference in Braunwald''s classification between the high thioredoxin and the low thioredoxin group on admission, as analyzed by the χ2 test with Yates''s correction (p<0.05). Moreover, there was a significant difference in incidence of recurrent anginal attacks at rest between the high thioredoxin and the low thioredoxin group after treatment, as analyzed by the χ2 test with Yates''s correction (p<0.001). The present study demonstrated that plasma thioredoxin levels are significantly increased in patients with unstable angina compared to those with stable exertional angina and chest pain syndrome. Thioredoxin levels were associated with recurrent myocardial ischemia in patients with unstable angina. [Copyright &y& Elsevier]

Discusses the identification of rpoBC genes encoding for β and β′ subunits of RNA polymerase in a deep-sea piezophilic bacterium, Shewanella violacea Strain DSS12. Purification of RNA polymerase from S. violacea cells; Characterization of S. violacea RNA polymerase; Structural analysis of the rpoBC genes of S. violacea; Overproduction and affinity purification of all subunit proteins for recombinant RNA polymerase.

We assessed serial changes in oxidative stress in patients with acute myocardial infarction by measuring urinary 8-hydroxy-2′-deoxyguanosine excretion. The urinary 8-hydroxy-2′-deoxyguanosine creatinine levels peaked at 4 hours after reperfusion therapy and the levels were higher in the cardiac event group than in the noncardiac event group before and after reperfusion. [Copyright &y& Elsevier]

The rpoE gene encoding an RNA polymerase s E subunit was isolated from a ?-phage library of the deep-sea piezophilic and psychrophilic bacterium Shewanella violacea strain DSS12. Structual analysis showed that the gene organization of the fragment containing S. violacearpoE was the l-aspartate oxidase-coding gene, rpoE, rseA, rseB and rseC in that order, the same as in the case of Photobacterium profundum SS9 and Escherichia coli K-12. The cloned gene, 576?bp in length, was found to encode a protein consisting of 192 amino acid residues with a molecular mass of 21,806?Da. Amino acid alignment of the RpoE protein showed that the functional domains responsible for DNA recognition, DNA melting, core binding, and RseA interaction were highly conserved. We purified hexahistidine-fused RpoE protein by constructing an overexpression plasmid. Core-binding analysis revealed that the cloned RpoE protein has the ability to bind with core RNA polymerase as a sigma factor. [ABSTRACT FROM AUTHOR]

RNA polymerase was purified from the piezophileShewanella violaceaDSS12, and the transcriptional activity after pressure treatment was compared with that of the mesophileEscherichia coli. Application of pressure at 100 MPa for 30 min reduced theE. coliRNA polymerase activity to 60% of the activity at atmospheric pressure, whereas theS. violaceaRNA polymerase maintained full activity, indicating that theS. violaceaRNA polymerase is more stable than itsE. colicounterpart. This result was supported by the analysis of the strength of subunit interactions of the enzyme from both species, using a high-pressure electrophoresis apparatus, which showed that a pressure of 140 MPa caused dissociation ofE. coliRNA polymerase but not that ofS. violaceaRNA polymerase. On the other hand, the core enzyme ofS. violaceaRNA polymerase, which lacked the s70 factor, was dissociated at 140 MPa. These results suggest that the s70 factor is required for stabilization ofS. violaceaRNA polymerase under high-pressure conditions. In this paper, we provide in vitro evidence for piezoadaptation at the transcriptional level, using purified RNA polymerase from cells ofS. violaceaandE. coli. [ABSTRACT FROM AUTHOR]

Introduction: The fibrinolytic system has a major role as a defense mechanism against thrombus formation. Net fibrinolytic activity in plasma reflects the balance between tissue-type plasminogen activator and plasminogen activator inhibitor (PAI). PAI is the main factor determining overall fibrinolytic activity. Materials and methods: We examined the effects of oral administration of vitamin E, an antioxidant, on fibrinolytic activity and oxidative stress in patients with coronary spastic angina. Forty patients with coronary spastic angina were randomly assigned into two treatment groups, either vitamin E group (α-tocopherol acetate, 400 mg/day) or placebo group by means of computerized system. PAI activity and thioredoxin, a marker of oxidative stress, levels were measured before and at the end of 1 month treatment. Results: Before treatment, the levels of PAI activity and thioredoxin were increased in patients with coronary spastic angina as compared with control subjects (n=17) (PAI activity levels: 13.6±1.4 vs. 7.6±2.2 IU/ml, p<0.05, thioredoxin levels: 22.8±1.7 vs. 16.0±1.4 ng/ml, p<0.05). In patients with coronary spastic angina, administration of vitamin E decreased both PAI activity and thioredoxin levels (PAI activity levels: 14.7±1.7 to 7.5±1.6 IU/ml, p<0.01, thioredoxin levels: 23.3±2.4 to 15.1±2.5 ng/ml, p<0.01), whereas placebo had no effect on these variables. Conclusions: Oral administration of vitamin E improved fibrinolytic activity and the improvement was associated with a decrease in oxidative stress. Administration of vitamin E is possible to be an effective adjunct therapy of coronary spasm in the absence of coronary atherosclerosis. [Copyright &y& Elsevier]

Objectives: We investigated the relationship between the urinary levels of biopyrrins and the severity of heart failure (HF).Background: Oxidative stress is evident in heart disease and contributes to the development of ventricular dysfunction in patients with HF. Biopyrrins, oxidative metabolites of bilirubin, have been discovered as potential markers of oxidative stress.Methods: We measured the levels of urinary biopyrrins and plasma B-type natriuretic peptide (BNP) in 94 patients with HF (59 men; mean age 65 years) and 47 control subjects (30 men; mean age 65 years). Urine and blood samples were taken after admission in all subjects. Further urine samples were obtained from 40 patients after treatment of HF.Results: The urinary biopyrrins/creatinine levels (μmol/g creatinine) were the highest in patients in New York Heart Association (NYHA) class III/IV (n = 26; 17.05 [range 7.85 to 42.91]). The urinary biopyrrins/creatinine levels in patients in NYHA class I (n = 35; 3.46 [range 2.60 to 5.42]) or II (n = 33; 5.39 [range 3.37 to 9.36]) were significantly higher than those in controls (2.38 [range 1.57 to 3.15]). There were significant differences in urinary biopyrrins/creatinine levels among each group. The treatment of HF significantly decreased both urinary biopyrrins/creatinine levels (from 7.43 [range 3.84 to 17.05] to 3.07 [range 2.21 to 5.71]) and NYHA class (from 2.5 ± 0.1 to 1.7 ± 0.1). Log biopyrrins/creatinine levels were positively correlated with log BNP levels (r = 0.650, p < 0.001).Conclusions: These results indicate that urinary biopyrrins levels are increased in patients with HF and are elevated in proportion to its severity. [Copyright &y& Elsevier]

We have cloned the rpoZ gene, encoding RNA polymerase ω protein, by PCR approach from the deep-sea piezophilic and psychrophilic bacterium, Shewanella violacea strain DSS12. The cloned gene, 285bp in length, was found to encode a protein consisting of 94 amino acid residues with a molecular mass of 10,327Da. Significant homology was evident comparing the RpoZ protein of S. violacea with that of Shewanella oneidensis (69% identity), Vibrio cholerae (65% identity), Escherichia coli K-12 (64% identity) and Haemophilus influenzae (61% identity). From the Northern blot analysis, S. violacea rpoZ gene was expressed constitutively under pressure conditions of 0.1, 30 and 50MPa. We constructed expression plasmid to overpmduce the RpoZ protein and transformed into E. coli JM109 as a host of overproduction. Upon induction, the recombinant protein encoded by plasmid pQrpoZ was overexpressed and purified using Ni2+ affinity column. [ABSTRACT FROM AUTHOR]

Although the blood pressure-lowering action of some long-acting calcium antagonists may last more than 24 h, some patients taking this drug still experience a morning surge, i.e. an early morning increase in blood pressure. We evaluated this morning surge in three hypertensive patients with morning surge after administration of antihypertensive drugs including nifedipine CR. Nifedipine CR is one of the once-a-day formulations of nifedipine, which after administration, results in two peaks in the plasma concentration of nifedipine. The first concentration peak of occurs 3 h after administration and the second around 12 h after intake. When the time of intake of nifedipine CR was changed from after breakfast to immediately after awakening, the morning surge was suppressed in these patients without the use of other drugs such as alpha- or beta-blockers. Administration of nifedipine CR immediately after awakening is one option that can be used to prevent morning surge as well as to control blood pressure throughout the day. [ABSTRACT FROM AUTHOR]

Although the blood pressure-lowering action of some long-acting calcium antagonists may last more than 24 h, some patients taking this drug still experience a morning surge, i.e. an early morning increase in blood pressure. We evaluated this morning surge in three hypertensive patients with morning surge after administration of antihypertensive drugs including nifedipine CR. Nifedipine CR is one of the once-a-day formulations of nifedipine, which after administration, results in two peaks in the plasma concentration of nifedipine. The first concentration peak of occurs 3 h after administration and the second around 12 h after intake. When the time of intake of nifedipine CR was changed from after breakfast to immediately after awakening, the morning surge was suppressed in these patients without the use of other drugs such as alpha- or beta-blockers. Administration of nifedipine CR immediately after awakening is one option that can be used to prevent morning surge as well as to control blood pressure throughout the day. [ABSTRACT FROM AUTHOR]

: ObjectivesThe aim of the present study was to investigate whether there is diurnal fluctuation in the endothelial function of patients with variant angina (VA).: BackgroundCoronary spasm is induced by acetylcholine and is promptly relieved by nitroglycerin. Thus, it is possible that endothelial dysfunction is involved in the pathogenesis of coronary spasm. Furthermore, the frequency of ischemic episodes is known to display diurnal variation.: MethodsFlow-mediated, endothelium-dependent vasodilation of the brachial arteries was measured in the early morning (6 am), afternoon (2 pm) and evening (8 pm) in 20 patients with VA (mean age 54.5 years; 10 men and 10 women) and in 20 control subjects (mean age 54.2 years; 10 men and 10 women). All patients underwent 24-h ambulatory electrocardiographic monitoring during the study.: ResultsFlow-mediated vasodilation in patients with VA was deteriorated by the early morning and improved by the afternoon (patients with VA at 8 pm vs. 6 am vs. 2 pm: 7.8 ± 2.1% (p < 0.01 vs. VA at 6 am) vs. 5.4 ± 2.3% vs. 8.8 ± 1.9% (p < 0.01 vs. VA at 6 am); control subjects: 9.5 ± 2.8% vs. 9.0 ± 2.2% vs. 9.9 ± 1.9%, respectively). The frequency of spontaneous ischemic episodes was highest from midnight to morning and was lowest from morning to late afternoon (4 pm to midnight: 7 episodes; midnight to 8 am: 25 episodes; 8 am to 4 pm: 3 episodes).: ConclusionsThere is diurnal fluctuation in endothelial function, which is associated with variation in the frequency of ischemic episodes. [Copyright &y& Elsevier]

Hypercholesterolemia impairs endothelial function. However, the production/release of nitric oxide from the hypercholesterolemic aorta is reported to be enhanced rather than impaired in animal studies. l-arginine improves endothelial function in hypercholesterolemic subjects. The goal of the present study was to investigate the effects of l-arginine on endothelial function and oxidative stress in hypercholesterolemic subjects. In 17 hypercholesterolemic male subjects (mean age 41.7 years, mean total cholesterol 264.3±5.9 mg/dl) and 17 age-matched healthy men as controls (mean total cholesterol 187.1±6.8 mg/dl), we measured flow-mediated endothelium-dependent vasodilation of the brachial artery during saline infusion and after saline plus l-arginine infusion (30 g for 1 h) with ultrasound technique. In addition, we measured the levels of thiobarbituric acid reactive substances (TBARS), as a marker of lipid peroxide. The flow-mediated vasodilation was lower and the TBARS concentration was higher in the hypercholesterolemic group than in the control group during the saline infusion. The addition of l-arginine increased flow-mediated vasodilation and decreased TBARS concentration in the hypercholesterolemic group (from 3.92±0.58 to 7.27±0.53% [P<0.01 by analysis of variance (ANOVA)], from 7.74±0.46 to 5.71±0.35 nmol/ml [P<0.01 by ANOVA], respectively), but not in the control group (from 7.74±0.40 to 8.21±0.47%, from 5.45±0.43 to 4.83±0.35 nmol/ml, respectively). The endothelial function is blunted, and the oxidative stress is increased in hypercholesterolemic subjects. l-arginine improves endothelial function with decreasing oxidative stress. The augmentation of nitric oxide production/release induced by l-arginine may act as an antioxidant, and contributes to the improvement of endothelial function in hypercholesterolemic subjects. [Copyright &y& Elsevier]

Background: An abundance of ovarian hormones is assumed to be a major contributor to the low incidence of ischemic heart disease in premenopausal women. However, the effects of ovarian hormones remain undetermined.Objective: To examine whether the variation in ovarian hormone levels throughout a menstrual cycle affects myocardial ischemia in women with variant angina.Design: Prospective, observational study.Setting: University medical center in Japan.Participants: 10 premenopausal women with variant angina.Measurements: Frequency of spontaneous ischemic episodes, flow-mediated dilation of brachial artery, and serum levels of estradiol and progesterone.Results: Frequency of ischemic episodes was highest from the end of the luteal phase to the beginning of the menstrual phase and was lowest in the follicular phase. Flow-mediated vasodilation and estradiol levels were lowest from the end of the luteal phase to the beginning of the menstrual phase and were highest in the follicular phase.Conclusions: In premenopausal women with variant angina, we documented a cyclic variation in endothelial function and the frequency of myocardial ischemia that was associated with the variation in estrogen levels. [ABSTRACT FROM AUTHOR]

Although echocardiographically determined left ventricular mass and geometry predict cardiovascular morbid events in patients with hypertension, the mechanisms underlying this relation are unclear. There is considerable evidence that endothelium-dependent vasodilation is impaired in patients with hypertension. Thus, endothelial dysfunction may contribute to the mechanism that causes cardiovascular morbid events. This study was designed to examine the relationship between left ventricular geometry and endothelial function in patients with hypertension. The percentage increase in brachial arterial diameter during reactive hyperemia was examined by a high-resolution ultrasound technique in 49 patients with hypertension and 64 normotensive subjects. Patients with hypertension had an impairment of the percentage increase in brachial arterial diameter during reactive hyperemia and an increase in thiobarbituric acid-reactive substances (TBARS) compared to normotensive subjects (percentage increase in diameter 5.6 ± 3.0 vs. 8.0 ± 2.5%, p < 0.001; TBARS levels 6.1 ± 1.3 vs. 5.3 ± 1.0 nmol/ml, p < 0.001). In patients with hypertension, there was a significant correlation between the left ventricular mass index and the percentage increase in brachial arterial diameter during reactive hyperemia (r = –0.583, p < 0.001), and the percentage increase in brachial arterial diameter during reactive hyperemia varied with the pattern of left ventricular geometry (normal ventricular geometry: 7.7 ± 2.6%; concentric remodeling: 5.2 ± 2.3%; eccentric hypertrophy: 4.2 ± 1.8%; concentric hypertrophy: 2.9 ± 2.6%). We conclude that (1) flow-mediated endothelium-dependent vasodilation in the brachial artery is impaired in patients with hypertension, (2) a relationship exists between the left ventricular mass index and flow-mediated endothelium-dependent vasodilation in the brachial artery in patients with hypertension and (3) increased oxidative stress may play a role in the endothelial dysfunction in patients with hypertension.Copyright © 2001 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Studies the effects of vitamin C on insulin resistance and endothelial dysfunction in smokers. Results of oral glucose tolerance tests; Influence of vitamin C on flow-mediated dilation of the brachian artery and plasms thiobarbituric acid-reactive substance; Induction of endothelium-dependent vasodilation by insulin.

Presents a study which examined the effects of vitamin C on insulin sensitivity and endothelial function in smokers. Account of related studies; Methods; Results and discussion.

Dobutamine stress echocardiography (DSE) is a useful and safe provocation test for myocardial ischemia. Until now, the test has been focused only on the organic lesion in the coronary artery, and positive DSE has indicated the presence of significant fixed coronary artery stenosis. The aim of the present study is to examine whether myocardial ischemia due to coronary spasm is induced by dobutamine. We performed DSE on 51 patients with coronary spastic angina but without significant fixed coronary artery stenosis. All patients had anginal attacks at rest with ST elevation on the electrocardiogram (variant angina). Coronary spasm was induced by intracoronary injection of acetylcholine, and no fixed coronary artery stenosis was documented on angiograms in all patients. DSE was performed with intravenous dobutamine infusion with an incremental doses of 5, 10, 20, 30, and 40 microg/kg/min every 5 minutes. Of the 51 patients, 7 patients showed asynergy with ST elevation. All 7 patients (13.7%) had chest pain during asynergy, and both chest pain and electrocardiographic changes were preceded by asynergy. These findings indicate that dobutamine can provoke coronary spasm in some patients with coronary spastic angina. When DSE is performed to evaluate coronary artery disease, not only fixed coronary stenosis, but also coronary spasm should be considered as a genesis of asynergy. [ABSTRACT FROM AUTHOR]

Examines the effect of vitamin C infusion on flow-mediated, endothelium-dependent vasodilation of the brachial artery in smokers and nonsmokers. Hemodynamic variables; Flow-dependent vasodilation; Levels of serum vitamin C and plasma thiobarbituric acid-reactive substances (TBARS); Correlation between serum level of vitamin C and flow-dependent vasodilation.

Examines the effects of vitamin C and cigarette smoking on endothelium-dependent vasodilation. Methodology used in study; Measurement of the serum levels of vitamin C and plasma levels of thiobarbituric acid-reactive substances (TBARS); Impact of vitamin C administration on impairment of flow-dependent vasodilation; Conclusion on endothelium-dependent vasodilation in the brachial arteries.

The article presents a study which identifies and characterize two alternative σ factors, σ E2 and σ E3 of RNA polymerase in the deep-sea piezophilic bacterium Shewanella violecea DSS12. The alignments of amino acid showed comparative conservation of domains for transcriptional functions compared with Escherichia coli σ E in both proteins. It notes that both proteins functions as σ factors as revealed by its core-binding analysis.

Abstract: The secretion of dehydroepiandrosterone sulfate (DHEAS) decreases with age, and the incidence of heart failure rises in the elderly population. We measured plasma DHEAS levels in 50 male patients (mean 66.7±9.1 years old) with congestive heart failure due to idiopathic dilated cardiomyopathy before and after treatment. The study included 50 age-matched control subjects with coronary spastic angina (mean 65.5±8.8 years old). DHEAS levels were significantly lower in patients with congestive heart failure than in controls (82.2±9.9 vs. 122.7±18.6 µg/dL, respectively, p <0.01), whereas there was no difference in cortisol levels between the 2 groups. After 3 months of treatment, NYHA functional class improved in all patients, and DHEAS levels increased (from 82.2±9.9 to 106.2±21.1 µg/dL, p <0.01). DHEAS levels vary according as heart failure condition in patients with idiopathic dilated cardiomyopathy. [Copyright &y& Elsevier]

Investigates the protein-DNA complex composed of NtrC protein and its target promoter DNA. Analysis of the protein-DNA interaction using capillary narrow-tube electrophoresis; Behavior of the complex under high-pressure conditions; Role of the complex in pressure-regulated transcription process.

We investigated whether the assessment of small platelet aggregates before percutaneous coronary intervention (PCI) could predict restenosis after PCI. This was a prospective cohort study that enrolled 189 consecutive patients who had coronary artery disease. In multiple logistic regression analysis, higher levels of preprocedural small platelet aggregates were independently associated with restenosis after PCI. Measurement of small platelet aggregates may serve as a useful clinical variable for stratifying patients who present for PCI. [Copyright &y& Elsevier]

Describes the reconstitution and characterization of bacterial enhancer-binding protein NtrC in a deep-sea piezophilic bacterium, Shewanella violacea. Recognition of the sequence for NtrC binding within the region upstream of the glnA operon; Expression of NtrC at a higher level under high pressure conditions.

Discusses the effect of medroxyprogesterone acetate plus (MPA) estradiol on endothelium-dependent vasodilation in postmenopausal women. Study methods and subjects; Effect of estrogen replacement therapy on lipid profiles and risk of cardiovascular events; Effect of estradiol therapy with or without MPA, on vascular reactivity.

Ritonavir (RTV), which is used in combination with nilmatrelvir (NMV) to treat coronavirus disease 2019 (COVID-19), inhibits cytochrome P450 (CYP) 3A, thereby increasing blood tacrolimus (TAC) levels through a drug–drug interaction (DDI). We experienced a case in which a DDI between the two drugs led to markedly increased blood TAC levels, resulting in vasospastic angina (VSA) and acute kidney injury (AKI). Rifampicin (RFP) was administered to induce CYP3A and promote TAC metabolism. A 60-year-old man with dermatomyositis who was taking 3 mg/day TAC contracted COVID-19. The patient started oral NMV/RTV therapy, and he was admitted to the hospital after 4 days because of chest pain and AKI. On day 5, his blood TAC level increased markedly to 119.8 ng/mL. RFP 600 mg was administered once daily for 3 days, and his blood TAC level decreased to the therapeutic range of 9.6 ng/mL on day 9, leading to AKI improvement. Transient complete atrioventricular block and nonsustained ventricular tachycardia were present during chest pain. In the coronary spasm provocation test, complete occlusion was observed in the right coronary artery, leading to a diagnosis of VSA. VSA and AKI are possible side effects of high blood TAC levels caused by DDI, and attention should be paid to cardiovascular side effects such as VSA and AKI associated with increased blood levels of TAC when it is used together with NMV/RTV. When blood levels of TAC increase, oral RFP can rapidly decrease TAC blood levels and potentially reduce its toxicity. [ABSTRACT FROM AUTHOR]