Revisit to diffraction anomalous fine structure.

The diffraction anomalous fine structure (DAFS) method that is a spectroscopic analysis combined with resonant X-ray diffraction enables the determination of the valence state and local structure of a selected element at a specific crystalline site and/or phase. This method has been improved by using a polycrystalline sample, channel-cut monochromator optics with an undulator synchrotron radiation source, an area detector and direct determination of resonant terms with a logarithmic dispersion relation. This study makes the DAFS method more convenient and saves a large amount of measurement time in comparison with the conventional DAFS method with a single crystal. The improved DAFS method has been applied to some model samples, Ni foil and Fe3O4 powder, to demonstrate the validity of the measurement and the analysis of the present DAFS method.

Chronic kidney disease categories and renal-cardiovascular outcomes in type 2 diabetes without prevalent cardiovascular disease: a prospective cohort study (JDDM25).

AIMS/HYPOTHESIS: In type 2 diabetic patients at low risk for cardiovascular disease (CVD), the relationship between the clinical course of nephropathy by stage of chronic kidney disease (CKD) and onset of CVD remains unclear. Clarification of this relationship is important for clinical decision-making for both low- and high-risk diabetic patients. METHODS: This 4 year prospective study enrolled 2,954 type 2 diabetic patients with no prevalent CVD, and serum creatinine <176.8 µmol/l. The risk for CVD onset (non-fatal and fatal CVD and stroke, and peripheral arterial disease) was assessed according to CKD stage categorised by urinary albumin-to-creatinine ratio (ACR; mg/mmol) and estimated GFR (eGFR; ml min(-1) 1.73 m(-2)). Association of progression from 'no CKD' stage (ACR <3.5 mg/mmol and eGFR ≥ 90 ml min(-1) 1.73 m(-2)) with risk for CVD onset was also evaluated. RESULTS: During follow-up (median 3.8 years), 89 CVD events occurred. Compared with patients with 'no CKD' as reference, those with ACR ≥ 35.0 mg/mmol with co-existing eGFR 60-89 ml min(-1) 1.73 m(-2) or <60 ml min(-1) 1.73 m(-2) showed increased risk for CVD onset, whereas those with eGFR ≥ 90 ml min(-1) 1.73 m(-2) did not. Those with ACR <3.5 mg/mmol and eGFR <60 ml min(-1) 1.73 m(-2) did not show any increased risk. Among patients with 'no CKD' stage at baseline, those who progressed to ACR ≥ 3.5 mg/mmol during follow-up showed an increased risk compared with those who did not, whereas those who progressed to eGFR <90 ml min(-1) 1.73 m(-2) did not have increased risk. CONCLUSIONS/INTERPRETATION: The risk for CVD was associated with progression of albuminuria stage rather than eGFR stage in type 2 diabetic patients at relatively low risk for CVD.

Comparison of the prognostic value of inflammation-based prognostic scores in patients with hepatocellular carcinoma.

BACKGROUND: Inflammation-based prognostic scores including the Glasgow Prognostic Score (GPS), neutrophil to lymphocyte ratio (NLR), and Prognostic Nutritional Index (PNI) are associated with survival in patients with hepatocellular carcinoma (HCC). The aim of this study was to investigate the prognostic value of these inflammation-based prognostic scores in patients with HCC. METHODS: In total, 150 patients with newly diagnosed HCC were prospectively evaluated. Patients were divided according to the GPS, modified GPS, NLR, platelet to lymphocyte ratio (PLR), Prognostic Index (PI), and PNI. The area under the receiver operating characteristics curve (AUC) was calculated to compare the predictive ability of each of the scoring systems. A univariate and multivariate analysis were performed to identify the clinicopathological variables associated with overall survival. RESULTS: The GPS consistently had a higher AUC value at 6 months (0.768), 12 months (0.787), and 24 months (0.758) in comparison with other inflammation-based prognostic scores. A multivariate analysis showed that the GPS was independently associated with overall survival. CONCLUSION: This study demonstrates that the GPS, an inflammation-based prognostic score, is an independent marker of poor prognosis in patients with HCC and is superior to the other inflammation-based prognostic scores in terms of prognostic ability.

Gender differences in maximum voluntary lip-closing force during lip pursing in healthy young adults.

The objectives of this study were to quantitatively evaluate the gender differences in the lip-closing force (LCF) generated during pursing-like lip-closing movement using a multidirectional LCF measurement system in healthy young adults. In 40 healthy subjects (20 women, 20 men; median age = 26·5 years, range = 22-41 years), LCF was recorded in eight directions during the performance of a voluntary pursing-like lip-closing task in four measurement sessions. The correlations between the total sum of the forces generated in all eight directions [total LCF (TLCF)] and each directional LCF (DLCF) and those between opposing DLCF were statistically analysed. The TLCF obtained from the highly reproducible measurements acquired in the four different sessions was normally distributed in both genders. The TLCF in men was significantly greater than that in women. Among the eight pairs of opposing DLCF, seven pairs of opposing DLCF showed significant correlations in men, while five pairs were significantly correlated in women. In men, no significant difference was observed between opposing DLCF; however, three pairs of opposing DLCF were significantly different in women. The present results quantitatively indicate that there are gender differences in the magnitude and directional specificity of the LCF produced during pursing-like lip-closing movement in healthy young adults.

Low incidence of cardiovascular events in Japanese patients with Type 2 diabetes in primary care settings: a prospective cohort study (JDDM 20).

AIMS: To investigate whether a reduced incidence of cardiovascular disease in Type 2 diabetes can be achieved in a newly recruited cohort following the recently advanced concept of multifactorial treatment and followed in primary care settings as compared with earlier cohorts. METHODS: A prospective study was performed in primary care settings at multiple clinics nationwide in the Japan Diabetes Clinical Data Management (JDDM) study group. Subjects were 2984 patients with Type 2 diabetes without prevalent cardiovascular disease. The main outcome measure was the first event of non-fatal or fatal coronary heart disease, ischaemic stroke or peripheral artery disease, and the incidence was compared with other representative cohorts. RESULTS: There were 90 cardiovascular events over 10,827 person-years of follow-up with a dropout rate of 6%. The incidences (per 1000 person-years, 95% confidence interval) of composite, coronary heart disease, ischaemic stroke and peripheral artery disease in the JDDM study were 8.3 (6.6-10.0), 4.4 (3.2-5.6), 3.1 (2.1-4.2), and 0.7 (0.2-1.2), respectively. Each incidence was lowest in the JDDM study compared with other cohorts (P < 0.01 vs. each cohort). In the JDDM study, significant variables predictive of the occurrence of a cardiovascular event were age, duration of diabetes, HbA(1c), HDL cholesterol and urinary albumin. CONCLUSION: The novel finding of low cardiovascular disease occurrence in this study may be conferred by the feasibility at primary care settings for providing patients with Type 2 diabetes with favourable control of blood glucose, blood pressure and lipids, coupled with unique ethnicity/country factors.

Active hexose correlated compound inhibits the expression of proinflammatory biomarker iNOS in hepatocytes.

BACKGROUND/AIMS: Excess production of nitric oxide (NO) by inducible nitric oxide synthase (iNOS) has been implicated as proinflammatory biomarker in liver injury. The application of active hexose correlated compound (AHCC) as a functional food in complementary and alternative medicine has increased. The possibility that AHCC might inhibit iNOS induction was investigated as a potential liver-protective effect. METHODS: Hepatocytes were isolated from rats by collagenase perfusion and cultured. Primary cultured hepatocytes were treated with interleukin-1ß in the presence or absence of AHCC-sugar fraction (AHCC-SF). RESULTS AND CONCLUSION: AHCC-SF inhibited the production of NO and reduced expressions of iNOS mRNA and its protein. AHCC-SF had no effects on either IκB degradation or nuclear factor-κB (NF-κB) activation. In contrast, AHCC-SF inhibited the upregulation of type I interleukin-1 receptor (IL-1RI) through the inhibition of Akt phosphorylation. Transfection experiments with iNOS promoter-luciferase constructs revealed that AHCC-SF reduced the levels of iNOS mRNA at both promoter transactivation and mRNA stabilization steps. AHCC-SF inhibited the expression of iNOS gene antisense transcript, which is involved in iNOS mRNA stabilization. These findings demonstrate that AHCC-SF suppresses iNOS gene expression through a IκB/NF-κB-independent but Akt/IL-1RI-dependent pathway, resulting in the reduction of NO production. AHCC-SF may have therapeutic potential for various liver injuries.

Regulatory relationship between tactile sensation at the vermilion of the lips and lip-closing force.

The objectives of this study were to identify the regulatory relationship between tactile sensation at the vermilion of the lips and the output of pursing-like lip-closing force (LCF), as measured by a multidirectional LCF measurement system. Thirty-six healthy subjects were divided into Anaesthesia and Vaseline groups. The tactile detection threshold (TDT) at six sites on the vermilion and the maximum voluntary LCFs in eight directions were recorded before and during partial or whole application of the agent and 6 h after whole application (Recovery). Five per cent lidocaine gel and Vaseline was applied to the vermilion in the Anaesthesia and Vaseline groups, respectively. These agents were applied to either the right part of the vermilion of the lower lip (Partial) or the whole vermilion (Whole). Partial application of 5% lidocaine gel significantly decreased the magnitudes of six of eight directional LCFs, while it only increased the TDT at one site. The subsequent whole application of 5% lidocaine gel did not affect the magnitude of the LCFs in five of these six directions although its application increased the TDTs at all sites. These reductions in LCF were reversed after recovery from surface anaesthesia. Vaseline application did not affect either TDT or LCF in any conditions. We concluded that tactile sensation at the vermilion of the lips is related to the output of LCF, without any particular directional specificity. The present results suggest the presence of a common synaptic drive ordering the pursing of the relevant muscles.

Reliability of novel multidirectional lip-closing force measurement system.

This study aimed to quantify the directional specificity of multidirectional lip-closing force (LCF) and evaluate the reliability of multidirectional LCF measurements made using a novel system. In fourteen healthy subjects (seven females, seven males, median age = 28 years), LCFs in eight directions and electromyograms (EMGs) from four parts of the orbicularis oris muscles (OOM) were recorded during voluntary pursing-like lip closure tasks. The quantitative reliability was assessed from repeated measurements of the LCFs in the eight directions and from summed values for all eight directions [total lip-closing force (TLCF)]. The intra- and inter-investigator reliabilities for TLCF were assessed by the interclass correlation of the measurements by the same investigator and two investigators, respectively. Lip-closing forces showed directional specificity in vertical, horizontal and oblique directions but those in oblique and horizontal directions were symmetrical bilaterally. The quantitative reliability of measurements was between 0·735 and 0·948 in the eight directions and that of TLCF was 0·934. Interclass correlations of intra- and inter-investigator reliabilities were 0·96 [lower limit of 95% confidence interval (95% LL), 0·87] and 0·96 (95% LL, 0·91), respectively. The intra- and inter-investigator differences of measurements were randomly distributed in the whole range of measurements. The 95% confidence intervals of these differences were significantly narrower than those of the limits of agreement (mean ± 1·96 s.d.). In 13 subjects, Pearson's correlation coefficients between LCF and EMGs from OOM were above 0·95. We conclude that this system has a reasonable quality and reliability for quantitative measurements of multidirectional LCF for evaluating lip functions.

Induction of in vitro differentiation of mouse embryonal carcinoma (F9) and erythroleukemia (MEL) cells by herbimycin A, an inhibitor of protein phosphorylation.

Herbimycin A is one of the benzenoid ansamycin antibiotics isolated from a culture of Streptomyces species (Omura, S., A. Nakagawa, and N. Sadakane. 1979. Tetrahedron Lett. 1979: 4323-4326). Recent studies have shown that the antibiotic not only inhibits the phosphorylation of p60src in Rous sarcoma virus- (RSV) infected cells, but also reverses the cellular phenotypes acquired by transfection with tyrosine kinase oncogenes (Uehara, Y., M. Hori, T. Takeuchi, and H. Umezawa. 1985. Jpn. J. Cancer Res. 76:672-675; Uehara, Y., M. Hori, T. Takeuchi, and H. Umezawa. 1986. Mol. Cell. Biol. 6: 2198-2206; Uehara, Y., Y. Murakami, S. Mizuno, and S. Kawai. 1988. Virology. 164: 294-298). These studies and other evidence indicate that the antibiotic inhibits a reaction(s) closely associated with the function of cellular tyrosine kinases. We have found that herbimycin A is an effective inducing agent capable of triggering differentiation in two typical mouse in vitro differentiation systems, which have been considered to be quite different in their mechanism of induction: endoderm differentiation of embryonal carcinoma (F9) cells and terminal erythroid differentiation of erythroleukemia (MEL) cells. The results suggest that there is a common step in the intracellular differentiation cascade which is, directly or indirectly, associated with phosphorylation at specific (tyrosine) residues of cellular proteins. The significance of this finding with respect to the molecular mechanism of in vitro differentiation is discussed.

Shift work is a risk factor for increased total cholesterol level: a 14-year prospective cohort study in 6886 male workers.

OBJECTIVES: The widespread adoption of 24 h continuous operations in a number of industries has resulted in an increase in shift work, which may influence lipid metabolism because of disturbed circadian rhythms, broken sleep and lifestyle problems. The objective of the present study was to assess the effect of shift work on serum total cholesterol as an index of lipid metabolism. METHODS: A 14-year prospective cohort study was conducted in day workers (n = 4079) and alternating shift workers (n = 2807) who received annual health check-ups between 1991 and 2005 in a Japanese steel company. The end-points were either a 20%, 25%, 30%, 35%, 40% or 45% increase in serum total cholesterol during the period of observation, compared with serum total cholesterol at entry to the study. The association between the job schedule type and increase in serum total cholesterol was investigated using multivariate pooled logistic regression analyses. The odds ratios for the effect of shift work were obtained after adjustment for a number of potential confounders. RESULTS: The significant odds ratios of alternating shift work (and 95% confidence intervals) were: >or=20%, 1.16 (1.07 to 1.26); >or=25%, 1.16 (1.05 to 1.28); >or=35%, 1.23 (1.05 to 1.43); >or=40%, 1.30 (1.07 to 1.58); and >or=45%, 1.28 (1.01 to 1.63) for serum total cholesterol. CONCLUSION: Generally the odds ratios of alternating shift work tended to be higher for stricter cut-points of relative increase in serum total cholesterol level. Our study in male Japanese workers revealed that alternating shift work adversely affected lipid metabolism.

Facial nerve motor-evoked potential monitoring during skull base surgery predicts facial nerve outcome.

OBJECTIVE: To determine whether monitoring facial nerve motor-evoked potentials (FNMEPs) elicited by transcranial electrical stimulation during skull base tumour surgery is useful for predicting facial nerve outcome. METHODS: This study analysed FNMEP findings in 26 patients with skull base tumours. Corkscrew electrodes positioned at C3 or C4 and Cz were used to deliver supramaximal stimuli (180-550 V). FNMEPs were recorded from the orbicularis oculi and oris muscles. The correlation between the final-to-baseline FNMEP ratio and postoperative facial nerve function was examined. RESULTS: Postoperative facial nerve function correlated significantly with the FNMEP ratios in the orbicularis oculi (p = 0.004) and orbicularis oris (p<0.001) muscles. An FNMEP ratio of <50% consistently predicted immediate postoperative facial palsy, although the degree of palsy differed among patients. All patients had satisfactory facial nerve function (House and Brackmann grades I and II) postoperatively if the FNMEP ratio remained at >50%. CONCLUSIONS: Intraoperative FNMEP monitoring can be useful for predicting facial nerve function after skull base surgery. This new method is a valuable adjunct to conventional facial nerve monitoring.

Reduced GFR and microalbuminuria are independently associated with prevalent cardiovascular disease in Type 2 diabetes: JDDM study 16.

AIMS: We investigated whether a reduced estimated glomerular filtration rate (eGFR) was associated with cardiovascular disease (CVD) prevalence, independent of the effect of microalbuminuria in patients with diabetes. METHODS: In a multicentre, large-scale cohort including 3002 Japanese patients with Type 2 diabetes without macroalbuminuria, the relationship of a reduced eGFR and microalbuminuria with CVD was investigated. RESULTS: Of those patients, 4.8% had a reduced eGFR and microalbuminuria, 12.7% had a reduced eGFR without microalbuminuria and 18.7% had microalbuminuria but normal eGFR. A reduced eGFR and microalbuminuria were each associated with a doubling of the prevalence of CVD. Compared with patients with no microalbuminuria/normal eGFR [odds ratio (OR) 1.0], the OR for CVD was significantly higher in those with a reduced eGFR without microalbuminuria (OR 1.97) and similarly higher in those with microalbuminuria without a reduced eGFR (OR 1.85). The OR was highest in those with both a reduced eGFR and microalbuminuria (OR 3.97, 95% confidence interval 2.55-6.20). The OR for CVD remained significant after adjustments for age, sex, hypertension, dyslipidaemia, smoking, body mass index, glycated haemoglobin and the duration of diabetes, and remained significant if the cut-off point for microalbuminuria was set at the median albumin : creatinine ratio (13.7 mg/g creatinine). In patients without microalbuminuria, a reduced eGFR was associated with CVD only in the older and male groups. CONCLUSION: A reduced eGFR and the presence of microalbuminuria were each associated with a near doubling of the prevalence of CVD, independently of traditional CVD risk factors and glycaemic control in patients with Type 2 diabetes.

Induction of the cytokine signal regulator SOCS3/CIS3 as a therapeutic strategy for treating inflammatory arthritis.

Immune and inflammatory systems are controlled by multiple cytokines, including ILs and INFs. These cytokines exert their biological functions through Janus tyrosine kinases and STAT transcription factors. One such cytokine, IL-6, has been proposed to contribute to the development of rheumatoid arthritis (RA). We found that STAT3 was strongly tyrosine phosphorylated in synovial tissue of RA patients, but not those with osteoarthritis. Blockade of the IL-6-gp130-JAK-STAT3-signaling pathway might therefore be beneficial in the treatment of RA. We show here that the mRNA for the endogenous cytokine signaling repressor CIS3/SOCS3 is abundantly expressed in RA patients. To determine whether CIS3 is effective in treating experimental arthritis, a recombinant adenovirus carrying the CIS3 cDNA was injected periarticularly into the ankle joints of mice with antigen-induced arthritis or collagen-induced arthritis (CIA). Periarticular injection of CIS3 adenovirus drastically reduced the severity of arthritis and joint swelling compared with control groups. CIS3 was more effective than a dominant-negative form of STAT3 in the CIA model. Thus, induction of CIS3 could represent a new approach for effective treatment of RA.

UV Irradiation induces an activity which stimulates Simian virus 40 rescue upon cell fusion.

UV irradiation of African green monkey cells greatly stimulated efficiency of simian virus 40 induction from simian virus 40-transformed Syrian hamster cells after cell fusion. The maximum inducing activity was observed at 15 to 20 h after irradiation but remained only transiently. The addition of cycloheximide after UV irradiation eliminated the stimulation of the activity.

Poly(ADP-ribose) polymerase inhibitors suppress UV-induced human immunodeficiency virus type 1 gene expression at the posttranscriptional level.

Gene expression of human immunodeficiency virus type 1 (HIV-1) is induced not only by trans activation mediated through a gene product (tat) encoded by the virus but also by treatment of virus-carrying cells with DNA-damaging agents such as UV light. Employing an artificially constructed DNA in which the chloramphenicol acetyltransferase gene was placed under the control of the HIV-1 long terminal repeat, we analyzed the induction process in HeLa cells and found that inhibitors of poly(ADP-ribose) polymerase suppressed UV-induced HIV-1 gene expression but not tat-mediated expression. We also found that suppression occurs at the posttranscriptional level. These results indicate that HIV-1 gene expression is activated by at least two different mechanisms, one of which involves poly-ADP ribosylation. A possible new role of poly-ADP ribosylation in the regulation of specific gene expression is also discussed.

Effect of mitochondrial protein synthesis inhibitors on erythroid differentiation of mouse erythroleukemia (Friend) cells.

When mouse erythroleukemia (MEL) cells were incubated in the presence of chloramphenicol (a specific inhibitor for mitochondrial protein synthesis) during the early stage of in vitro erythroid differentiation, the number of induced erythroid cells was greatly reduced. By use of cell fusion between two genetically marked MEL cells, this finding was further investigated. We found that the drug, along with other agents which inhibit mitochondrial protein synthesis, blocked the induction and turnover of the DMSO-inducible intracellular-erythroid-inducing activity (differentiation-inducing factor II) in a manner similar to that of cycloheximide, an inhibitor for nuclear protein synthesis. The inhibitory effect was confirmed by directly assaying differentiation-inducing factor II in the cell extracts. These results strongly suggest that mitochondrial protein synthesis is closely associated with in vitro erythroid differentiation of MEL cells.

Tumor promoter 12-O-tetradecanoylphorbol 13-acetate stimulates simian virus 40 induction by DNA-damaging agents and tumor initiators.

Simian virus 40 (SV40)-transformed Syrian hamster kidney cells produce infectious SV40 virus particles after treatments which damage DNA, such as UV irradiation or mitomycin C treatment. We have found that the induction of SV40 by DNA-damaging agents is greatly stimulated when a typical tumor promoter, 12-O-tetradecanoylphorbol 13-acetate (TPA), is present in the medium. Phorbol, which has a molecular structure similar to TPA but does not have any tumor-promoting activity, showed no such stimulatory effect on SV40 induction. This apparent synergistic effect of DNA-damaging agents and tumor promoter (TPA) was more pronounced when a tumor initiator, benzo [a]pyrene or 2-acetamido-fluorene, was combined with TPA. The effect of TPA on UV-triggered SV40 induction was greatly influenced by the timing of TPA addition to the culture medium, which was most efficient when addition of TPA was 5 to 20 h before UV irradiation. The effect of TPA, however, was not observed in SV40 rescue from hamster cells by cell fusion with permissive monkey (C7) cells.