[Correlation of the Blood Test Results Obtained Between Assays Using Microliter-Scale Fingertip Blood Samples Collected with a Novel Blood Collection Device and Conventional Venous Blood Assays].

In this study, we investigated the correlations of values of biochemical and hematological tests obtained using microliter-scale fingertip blood samples collected with a newly developed blood collection device with those using conventional venous blood. Eighty volunteer subjects were enrolled in the study. Blood sam- ples were drawn from the fingertip of the ring finger by a single puncture and 60 µL of each sample was promptly and accurately aspirated into a blood collection chip. Then the chip was tightly sealed with chip container and was shaken to mix the contents without dispersing. For biochemical tests except of HbAlc, blood was collected without anticoagulant and centrifuged to obtain 15 µL of serum which was then diluted with 190 pL of physiological saline for the assay. For hematological tests and HbAlc, the sample was as- sayed with blood collected using EDTA-2K. As a result, a good correlation of the test values was obtained between the assay with fingertip blood and that with venous blood. The correlation coefficients were found to be 0.97 in TG, T-CHO, HDL-C, LDL-C, GLU, ALT, γ-GTP, UA, BUN and HbAlc and ≥ 0.95 for WBC, RBC, Hgb, and Hct. These results suggest that our microliter-scale blood testing can be comparable to the assay using venous blood and may be useful as a rapid and simple test for determination of basic clinical tests near the reference intervals. [Original].

[Current status of ISO 15189 accreditation system].

The Japan Accreditation Board (JAB) mainly involves the ISO 15189 accreditation system with support from the Japanese Committee for Clinical Laboratory Standards (JCCLS). The currently available procedure to obtain accreditation is as below. Firstly, it is necessary for applicants to prepare ISO 15189 and related documents in each laboratory. Then a JAB assessor will conduct a preliminary assessment to check if the applicant is ready to be accredited. Subsequently, a team consisting of one to five JAB assessors and/or technical experts will conduct the initial assessment, usually for two days. Finally, the team will make a recommendation to the JAB Accreditation Committee for Medical Laboratory on its evaluation for accreditation. If the Accreditation Committee approves the recommendation of the assessment team, the applicant will be granted accreditation and issued with a certificate of accreditation. According to EU data in February 2011, about 1,300 medical laboratories obtained the ISO 15189 accreditation. The numbers of accredited laboratories are 482 in Germany, 276 in England, 209 in France, 100 in Czechoslovakia etc. Similarly, the data for the Asia-Pacific region in June 2011 showed that the numbers of accredited laboratories are 638 in Australia, 287 in India, 220 in Canada, 160 in Taiwan etc. Although 59 laboratories are accredited in Japan, the ISO 15189 accreditation is not so widespread compared with other countries. It is now expected that the government and/or related bodies will have sufficient understanding of this accreditation system to further its development in Japan. [Rinsho Byori 60: 653-659, 2012]

Correlation of the Blood Test Results Obtained between Assays Using Microliter-scale Fingertip Blood Samples Collected with a Novel Blood Collection Device and Conventional Venous Blood Assays: a Secondary Publication in English.

In this study, we investigated the correlations between biochemical and hematological test results obtained using microliter-scale fingertip blood samples collected with a newly developed blood collection device and those obtained using conventional venous blood. Eighty volunteer subjects were enrolled in this study. Blood samples were drawn from the fingertip of the ring finger by a single puncture, and 60-µL samples were promptly and accurately aspirated into a blood collection chip. Then the chip was tightly sealed in a chip container and was shaken to mix the contents without dispersion. For biochemical tests other than that for HbA1c, blood was collected without anticoagulant and centrifuged to obtain 15 µL of serum which was then diluted with 190 µL of physiological saline for the assay. For hematological tests and the test for HbA1c, the sample was assayed with blood collected using EDTA-2 K. Good correlations were obtained between the test results of the assay using fingertip blood and that using venous blood. The correlation coefficients were ≥0.97 for TG, T-CHO, HDL-C, LDL-C, GLU, ALT, γ-GTP, UA, BUN, and HbA1c and ≥0.95 for WBC, RBC, Hgb, and Hct. These results suggest that our microliter-scale blood testing system is comparable to assays using venous blood and may be useful as a rapid and simple test to determine basic clinical parameters that are close to the reference intervals.

A561C polymorphism of E-selectin is associated with ischemic cerebrovascular disease in the Japanese population without diabetes mellitus and hypercholesterolemia.

E-selectin, which is a member of the selectin superfamily of adhesion molecules, contributes to the leukocyte-endothelial cell attachments and is involved in the pathogenesis of thrombovascular diseases as a consequence. We investigated the A561C mutation in the E-selectin gene in 235 Japanese patients with ischemic cerebrovascular disease (CVD) and 301 age- and sex-matched healthy controls. Excluding the subjects with diabetes mellitus and hypercholesterolemia, the AC genotype frequencies of patients with ischemic CVD were higher than those of controls: 12.7% vs. 5.8% (P=0.04). Our results show that E-selectin gene polymorphisms represent an increased risk for ischemic CVD in the Japanese population without diabetes mellitus and hypercholesterolemia.

G501C polymorphism of oxidized LDL receptor gene (OLR1) and ischemic stroke.

The human lectin-like oxidized low-density lipoprotein receptor 1 (OLR1/LOX-1) is the major endothelial scavenger receptor against oxidized low-density lipoprotein (Ox-LDL), which has been implicated in the pathogenesis of atherosclerosis. We investigated the G501C mutation in the OLR1 gene in 235 Japanese patients with ischemic cerebrovascular disease (CVD) and 274 age- and sex-matched healthy controls using single nucleotide primer extension analysis (SNuPe). There was no significant difference in the polymorphism between patients with ischemic CVD and controls (GC+CC versus GG, p=0.48). The C allele was not significantly different between the patients and controls (C versus G, p=0.91). Our results show that the OLR1 gene polymorphism has little effect on an increased risk for ischemic CVD in the Japanese population.

Gorog Thrombosis Test: analysis of factors influencing occlusive thrombus formation.

We used the Gorog Thrombosis Test to analyze the factors influencing the occlusion time, which represents platelet activation and subsequent occlusive thrombus formation, in 132 healthy Japanese volunteers (116 men, 16 women; mean age, 45.0 +/- 12.0 years). The Gorog Thrombosis Test was designed to evaluate platelet aggregation and thrombolytic activity under a high shear stress condition (175 dynes/cm) in a native blood sample in vitro. The mean +/- SD occlusion time was 154.8 +/- 64.7 s (men, 153.4 +/- 64.2 s and women, 165.4 +/- 56.5 s). The occlusion time was inversely correlated with von Willebrand factor ristocetin cofactor activity (VWF:Rco) (r = -0.242, P = 0.0055) and von Willebrand factor antigen (r = -0.230, P = 0.0080). The mean occlusion time in the group with VWF:Rco of at least 170% (137 s) was significantly shorter than that in the group with VWF:Rco less than 170% (156 s, P < 0.05). Platelet counts, other coagulation markers and smoking showed no significant correlations with occlusion time. Red blood cells (r = -0.177, P = 0.0365), hemoglobin (r = -0.191, P = 0.0245) and hematocrit (r = -0.182, P = 0.0329) also showed inverse correlations with the occlusion time. This report is the first to clearly demonstrate the role of von Willebrand factor in the formation of occlusive thrombi in the Gorog Thrombosis Test.

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (2004). III. Secular changes in susceptibility].

The bacteria (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella spp. and Pseudomonas aeruginosa) isolated from patients diagnosed as urinary tract infections (UTIs) in 14 institutions in Japan were collected between August 2004 and July 2005. The susceptibilities of these bacteria to various antimicrobial agents were measured. The bacteria were divided into 2 groups consisting of uncomplicated UTIs and complicated UTIs (with and without indwelling catheter) based on their isolation origins. The results were compared with those obtained between 1995 and 2003. The drug sensitivity of S. aureus in this year was similar to those in up to the previous year and S. aureus showed the best susceptibility to vancomycin (VCM) and arbekacin (ABK). The drug sensitivity of E. faecalis in this year also was similar to those in up to the previous year. The susceptibility of E. coli to cephems in this year was generally good and was similar to those in up to the previous year. MIC90 of cefozopran (CZOP) was the most stable and 0.125 microg/mL or less since 1995. The susceptibility of E. coli to cefpirome (CPR) and cefotiam (CTM) also was good but to cefaclor (CCL), cefixime (CFIX), and cefpodoxime (CPDX) was largely decreased in complicated UTI groups. The sensitivity of E. coli to carbapenems also was good but to carumonam (CRMN) tended to decrease. The susceptibility of E. coli to quinolones, however, has largely changed and has decreased since 2003 in uncomplicated UTIs and 2000 in complicated UTIs. That was suggested the development of the resistance to the drug. The susceptibility of Klebsiella spp. to cefazolin (CEZ), CTM, CCL, CPDX, and cefditoren (CDTR) decreased in the previous year and recovered to the year before the previous year in this year. The susceptibility of Klebsiella spp. to other cephems was stable since 1995, especially against CZOP, the highest sensitivity (MIC90: < or = 0.125 microg/mL) was maintained. The susceptibility of Klebsiella spp. to carbapenems and CRMN also was good. The susceptibility of Klebsiella spp. to aminoglycosides was lower than to CZOP but was stable since 1995. The susceptibility of P. aeruginosa was generally low and has largely changed against the majority of the agents since 1995. The susceptibility of P. aeruginosa isolated from uncomplicated UTIs has largely changed against ceftazidime (CAZ), cefsulodin (CFS), CZOP, imipenem (IPM), meropenem (MEPM), aztreonam (AZT), CRMN, gentamicin (GM), and tobramycin (TOB). The susceptibility of P. aeruginosa isolated from complicated UTIs has largely changed against CSF, CZOP, MEPM, GM, and ciprofloxacin (CPFX). The susceptibility of P. aeruginosa isolated from complicated UTIs has been stable against amikacin (AMK). For annual changes in MIC50, TOB and IPM had a relatively stable and high activity (MIC50: 0.5-2 microg/mL).

[In-vitro susceptibilites to levofloxacin and various antibacterial agents of 18,639 clinical isolates obtained from 77 centers in 2004].

A total of 18,639 clinical isolates in 19 species collected from 77 centers during 2004 in Japan were tested for their susceptibility to fluoroquinolones (FQs) and other selected antibiotics. The common respiratory pathogens, Streptococcus pyogenes, Streptococcus pneumoniae, Moraxella catarrhalis and Haemophilus influenzae showed a high susceptible rate against FQs. The isolation rate of beta lactamase non-producing ampicillin-resistant H. influenzae was approximately three times as large as those of western countries. Most strains of Enterobacteriaceae were also susceptible to FQs. The resistance rate of Escherichia coli against FQs has however been rapidly increasing so far as we surveyed since 1994. The FQs-resistant rate in methicillin-resistant Staphylococcus aureus (MRSA) showed approximately 90% except for 36%. of sitafloxacin while FQs-resistant rate in methicillin-susceptible S. aureus (MSSA) was around 5%. The FQs-resistant rate of methicillin-resistant coagulase negative Staphylococci (MRCNS) was also higher than that of methicillin-susceptible coagulase negative Staphylococci (MSCNS), however, it was lower than that of MRSA. In Pseudomonas aeruginosa clinical isolates, 32-34% from UTI and 15-19% of from RTI was resistant to FQs. Acinetobacter spp. showed a high susceptibility to FQs. Although FQs-resistant Neisseria gonorrhoeae have not been increased in western countries, it is remarkably high in Japan. In this survey, isolates of approximately 85% was resistant to FQs.

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (2004). I. Susceptibility distribution].

The bacterial strains isolated from 490 patients diagnosed as having urinary tract infections (UTIs) in 14 institutions in Japan were collected between August 2004 and July 2005. The susceptibilities of them to many kinds of antimicrobial agents were measured. Of them, 577 strains were estimated as causative bacteria and used for the measurement. The strains consisted of 156 gram-positive bacterial strains (27.0%) and 421 gram-negative bacterial strains (73.0%). Against Staphylococcus aureus, arbekacin (ABK), vancomycin (VCM) showed the strongest activity and prevented the growth of all strains with 2 microg/mL. Against Enterococcus faecalis, ampicillin (ABPC) and VCM showed a strong antibacterial activity. The antibacterial activity of cephems to Escherichia coli was generally good, and especially cefozopran (CZOP) and cefpirome (CPR) showed the strongest activity (MIC90: < or = 125 microg/mL). Quinolone resistant E. coli [MIC of ciprofloxacin (CPFX): > or = 4 microg/mL] was detected at frequency of 18.8%, which was higher than that in the last year. Against Klebsiella pneumoniae, CZOP, meropenem (MEPM), and carumonam (CRMN) showed the strongest activity and prevented the growth of all strains with 0.125 microg/mL or less. The antibacterial activity of the other cephems was relatively good, and decrease in their activity observed in the last year study was not recognized. Against Serratia marcescens, imipenem (IPM) and gentamicin (GM) had the strongest antibacterial activity. Against Proteus mirabilis, CRMN showed the strongest activity and prevented the growth of all strains with 0.125 microg/mL or less. MEPM prevented the growth of all strains with 0.25 microg/mL. Next, cefmenoxime (CMX), ceftazidime (CAZ), CZOP, cefixime (CFIX), cefpodoxime (CPDX), and cefditoren (CDTR) showed a strong activity. The antibacterial activity of the drugs to Pseudomonas aeruginosa was generally low, and MIC90 of all the drugs was ranged from 32 to > 128 microg/mL except IPM and MEPM having 16 microg/mL. The antibacterial activities of CZOP and CAZ were considered to be relatively good on MIC50 comparison (MIC50: 2 microg/mL).

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (2004) II. Background of patients].

Six hundred six bacterial strains isolated from 490 patients diagnosed as having urinary tract infections (UTIs) in 14 institutions in Japan were collected between August 2004 and July 2005. The frequency of bacteria isolation stratified with patient clinical background was compared. The clinical background investigated included sex, age, type of infections, timing of antibiotics administration, and presence or absence of surgery affecting a decrease in defense against infection. The bacterial strains were stratified with the age and sex of the patients and the types of infections. In males, the number of patients aged less than 60 years was few and the complicated UTIs without indwelling catheter was observed most frequently. In females, the number of patients aged less than 60 years was comparatively more than in males. In all of ages except 0-19 and > or = 80 years, the ratio of the uncomplicated UTIs was high, accounting for 44.1-90.0% of all types of infections. In the present time, the bacteria most frequently isolated were Escherichia coli. Pseudomonas aeruginosa and Enterococcus faecalis also were relatively frequently isolated. E. coli most frequently isolated with the uncomplicated UTIs and P. aeruginosa and E. faecalis most frequently isolated with the complicated UTIs. With respect to the relation of these results to the age of the patients, in the uncomplicated UTIs, the isolation frequency of E. coli was the highest in all age groups except 0-19 years, accounting for 50% or higher. In the complicated UTIs without indwelling catheter, the isolation frequency of E. coli tended to be high in all age groups. In the complicated UTIs with indwelling catheter, P. aeruginosa were more frequently isolated. In comparison of causative bacteria in UTIs between before and after the administration of antibiotics, P. aeruginosa increased after the administration in any types of UTIs. In comparison of causative bacteria in UTIs with or without surgery, E. coli was more frequently isolated in the patients without surgery, while P. aeruginosa and E. faecalis were more frequently isolated in the patients with surgery in any UTIs.

Salt intake affects the relation between hypertension and the T-786C polymorphism in the endothelial nitric oxide synthase gene.

BACKGROUND: Recent genetic studies have shown an association between the T-786C polymorphism in the endothelial NO synthase gene and coronary artery diseases, but any possible association with hypertension has been controversial. Other studies indicate the effect of restricting salt intake differ depending on individual salt-sensitivity, and the mixture of different sensitivity in study subjects may obscure the results. The objective of this study was to investigate the gene-environment interaction between the salt intake and this polymorphism. METHODS: We genotyped 281 healthy men after excluding 37 men on hypertensive therapy (mean age 44.8 +/- 11.9 years) for the mutation, and evaluated their daily salt intake using a validated food frequency questionnaire. RESULTS: A quartile classification of salt intake revealed that the blood pressure of subjects with the mutation was significantly higher than that of subjects without the mutation, but only in the 4th quartile (the highest intake group). A multiple logistic regression analysis also showed that the presence of this mutation increased the risk of hypertension only in the 4th quartile (adjusted odds ratio = 6.38, P = .025). CONCLUSIONS: The presence of this mutation alone does not significantly increase the risk of hypertension. However, high salt intake interacts with the mutation and leads to a significant increase in the risk of hypertension. The T-786C mutation warrants being considered a candidate for further study with the aim of tailor-made hypertension prevention.

T280M and V249I polymorphisms of fractalkine receptor CX3CR1 and ischemic cerebrovascular disease.

The contribution to atherosclerosis of two CX3CR1 single nucleotide polymorphisms, V249I and T280M has been recently reported. The atherosclerosis of intracranial vessels is thought to be the major pathological mechanism of ischemic stroke. In this study, we investigated the risk of ischemic stroke associated with fractalkine receptor CX3CR1 polymorphisms. We investigated the T280M and V249I mutations in the CX3CR1 gene in 235 Japanese patients with ischemic cerebrovascular disease (CVD) and 306 age- and sex-matched healthy controls. Polymerase chain reaction and restriction fragment length polymorphism were used for genotyping. There was no significant difference in both polymorphisms between patients with ischemic CVD and controls (VV versus II+VI, p=0.83; TT versus MM+TM, p=0.66). The I and M allele frequencies were not significantly different between CVD patients and controls: odds ratio (OR)=0.89 (95% confidence interval (CI)=0.50-1.60, p=0.70) and OR=1.19 (95% CI=0.71-2.00, p=0.51), respectively. We found eight of nine possible combined genotypes, including a new haplotype V249-M280, in Japanese. Our results show that these CX3CR1 gene polymorphisms are not associated with an increased risk for ischemic CVD in the Japanese population.

Factor XII Shizuoka, a novel mutation (Ala392Thr) identified and characterized in a patient with congenital coagulation factor XII deficiency.

We identified a novel mutation (Ala392Thr) in the factor XII (FXII) gene of a patient with congenital FXII deficiency, designated Factor XII Shizuoka. The proband was an asymptomatic 63-year-old Japanese male with an abnormal coagulation test, discovered by chance during preoperative testing. The FXII activity was under 3% and antigen level was under 10%. Sequence analysis of the proband's FXII gene revealed a homozygous nucleotide substitution G to A in exon 10, resulting in the amino acid substitution Ala392 to Thr in the catalytic domain. We constructed the mutant FXII cDNA in an expression plasmid vector and transfected it into Chinese hamster ovary (CHO) cells. The recombinant wild-type FXII antigen was detected in the culture medium by immunoprecipitation assay, but the mutant FXII (A392T) was not observed. Both the wild-type FXII and A392T cell lysates, however, contained equivalent levels of FXII antigen and FXII mRNA, as estimated by Western blotting and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), respectively. These findings suggest that the Ala392 to Thr substitution impairs intracellular protein processing and causes a cross-reacting material -negative FXII deficiency.

Inhibition of cellular adhesion in human umbilical vein endothelial cells by NF-kappaB inhibitor DHMEQ under flow.

We previously designed and synthesized DHMEQ as an inhibitor of NF-kappaB. In the present study, we looked into the effect of DHMEQ on the cell adhesion in human umbilical vein endothelial cells (HUVEC) under flow. We used freshly prepared HUVEC and human mononuclear cells throughout the experiment. DHMEQ inhibited TNF-alpha-, IL-1beta-, and LPS-induced NF-kappaB activation in HUVEC. It also inhibited TNF-alpha-induced expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin. DHMEQ also inhibited TNF-alpha-induced mononuclear cell-HUVEC adhesion. The effect of DHMEQ was more prominent when the cells were under shear stress. DHMEQ inhibited the adhesion between HUVEC and HT-29 colon cancer cells more clearly under the flow condition than under the static condition of the culture medium. These results suggest that DHMEQ, being a unique inhibitor of NF-kappaB, may be effective in suppressing atherosclerosis and metastasis by inhibiting the expression of adhesion molecules.

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (2003). I. Susceptibility distribution].

The bacterial strains isolated from 565 patients diagnosed as having urinary tract infections (UTIs) in 14 institutions in Japan were collected between August 2003 and July 2004. The susceptibilities of them to many kinds of antimicrobial agents were investigated. Of them, 701 strains were estimated as prophlogistic bacteria and used for the investigation. The strains consisted of 258 Gram-positive bacterial strains (36.8%) and 443 Gram-negative bacterial strains (63.2%). Against Staphylococcus aureus, vancomycin (VCM) showed the strongest activity and prevented the growth of all strains with 2 microg/mL. Against Streptococcus agalactiae, ampicillin (ABPC), cefozopran (CZOP), imipenem (IPM), and clarithromycin (CAM) showed a strong activity and the MIC90 was 0.125 microg/mL or less. Against Enterococcus faecalis, VCM, ABPC, and IPM showed a strong antibacterial activity. The antibacterial activity of cephems to Escherichia coli was generally good, and especially CZOP and cefpirome (CPR) showed the strongest activity (MIC90: < or = 0.125 microg/mL). Quinolone resistant E. coli [MIC of ciprofloxacin (CPFX): > or =4 microg/mL] was detected at frequency of 15.7%, which was higher than that in the last year. Against Klebsiella pneumoniae, meropenem (MEPM) showed the strongest activity and next, the antibacterial activity of CRMN and CZOP was good. The antibacterial activity of the other cephems, however, significantly decreased, compared with that evaluated in last year. Against Serratia marcescens, MEPM had the strongest antibacterial activity. Against Proteus mirabilis, MEPM and CRMN showed the strongest activity and prevented the growth of all strains with 0.125 microg/mL or less. Nest, cefmenoxime (CMX), ceftazidime (CAZ), cefixime (CFIX), cefpodoxime (CPDX), CPR, CZOP, and cefditoren (CDTR) showed a strong activity. The antibacterial activity of the drugs to Pseudomonas aeruginosa was generally low, and MIC90 of all the drugs was ranged from 32 to < or = 256 microg/mL except IPM and amikacin (AMK) having 16 microg/mL. The antibacterial activity of CZOP was relatively good (MIC50: 2 microg/mL).

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (2003). II. Background of patients].

Seven hundred and nineteen bacterial strains isolated from 565 patients diagnosed as having urinary tract infections (UTIs) in 14 institutions in Japan were collected between August 2002 and July 2003. The frequency of bacteria isolation divided with patient clinical background was compared. The clinical background investigated included sex, age, type of infections, timing of antibiotics administration, and presence or absence of surgery affecting a decrease in defense against infection. The bacterial strains were divided with the age and sex of the patients and the types of infections. In males, the number of patients aged less than 50 years was few and the complicated UTIs without indwelling catheter was observed most frequently. Number of patients aged 20-39 years was greater in female than male. In all of ages except 0-9 and 70-79 years, the ratio of the uncomplicated UTIs was high, accounting for 44.4-91.7% of all types of infections. In the present time, the bacteria most frequently isolated were Escherichia coli, Pseudomonas aeruginosa and Enterococcus faecalis also were relatively frequently isolated. E. coli was most frequently isolated from the uncomplicated UTIs, and P. aeruginosa and E. faecalis were frequently isolated from the complicated UTIs with indwelling catheter. With respect to the relation of these results to the age of the patients, in the uncomplicated UTIs, the isolation frequency of E. coli was the highest in all age groups, accounting for 40% or higher. In the complicated UTIs without indwelling catheter, the isolation frequency of E. coli decreased with aging of the patients but still was the highest in all age groups. In the complicated UTIs with indwelling catheter, the isolation frequency of E. coli was lower than in the uncomplicated UTIs in all age groups and P. aeruginosa and E. faecalis were more frequently isolated. In comparison of causative bacteria in UTIs between before and after the administration of antibiotics, P. aeruginosa increased after the administration in any types of UTIs. In comparison of causative bacteria in UTIs with or without surgery, E. coli was more frequently isolated in the patients without surgery, while P. aeruginosa and E. faecalis were more frequently isolated in the patients with surgery in any UTIs.

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (2003). III. Secular changes in susceptibility].

The bacteria (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella spp. and Pseudomonas aeruginosa) isolated from 565 patients diagnosed as urinary tract infections (UTIs) in 14 institutions in Japan were collected between August 2003 and July 2004. The susceptibilities of these bacteria to various antimicrobial agents were examined. The bacteria were divided into 2 groups consisting of uncomplicated UTIs and complicated UTIs (with and without indwelling catheter) based on their isolation origins. The results were compared with those obtained between 1994 and 2002. The drug sensitivity of S. aureus in this year was similar to those in up to the previous years and S. aureus showed the best susceptibility to vancomycin. The drug sensitivity of E. faecalis in this year also was similar to those in up to the previous years. The drug sensitivity of E. coli in this year was generally good except penicillins and was similar to those in up to the previous years. Among cephems, cefozopran (CZOP) and cefpirome (CPR) showed the highest potency activity (MIC90: < or = 0.125 microg/mL). An antibacterial activity of cefotiam (CTM) was stable for 10 years and was fine (MIC0: < or = 0.5 microg/mL). The sensitivity of E. coli to carbapenems and carumonam (CRMN) also was good like to CZOP. The sensitivity of the complicated UTIs group to quinolones, however, has decreased after 2000 and it was suggested that the resistance to the drug has developed. Kiebsiella spp. showed a decrease in the susceptibility to some of cephems. The drugs indicating a big decrease in the sensitivity were cefazolin, CTM, cefaclor, and cefpodoxime. Imipenem, carbapenems, also indicated a decrease in the sensitivity. The susceptibility of the strain to the other drugs was similar to that in up to the previous years. Among them, CZOP maintained good susceptibility (MIC90: > or = 0.125 microg/mL against uncomplicated UTIs, 0.25 microg/mL against complicated UTIs) like meropenem. The drug sensitivity of P. aeruginosa was generally low and was not much different from that in up to the previous years.

[In vitro susceptibilites to levofloxacin and various antibacterial agents of 11,475 clinical isolates obtained from 52 centers in 2002].

The susceptibilities of bacteria to fluoroquinolones (FQs), especially levofloxacin, and other antimicrobial agents were investigated using 11,475 clinical isolates collected in Japan during 2002. Methicillin susceptible staphylococci, Streptococcus pyogenes, Streptococcus pneumoniae, Moraxella catarrhalis, the family of Enterobactericeae, Haemophilus influenzae and Acinetobacter spp. exhibited stable and high susceptibilities to FQs. The rate of FQs-resistant MRSA was 80 approximately 90%, being markedly higher than that of FQs-resistant MSSA. The FQs-resistance rate of MRCNS was also higher than that of MSCNS, however, it was lower than that of MRSA. No FQs-resistant clinical isolates of Salmonella spp. were detected in any of the surveys. Thirteen of Escherichai coli 696 isolates, 8 of Klebsiella pneumoniae 630 isolates and 33 of Proteus mirabilis 373 isolates produced extended-spectrum beta-lactamase (ESBL), furthermore 6 of 13 in E. coli, 1 of 8 in K. pneumoniae and 14 of 31 ESBL-producing isolates, and in P. mirabilis were FQs resistant. Attention should be focused in the future on the emergence of ESBL in relation to FQs resistance. The rate of FQs-resistant P. aeruginosa isolated from urinary tract infection (UTI) was 40 approximately 60%, while 15 approximately 25% of isolates from respiratory tract infection (RTI) were resistant. IMP-1 type metallo beta-lactamase producing organisms were found in 49 of P. aeruginosa 1,095 isolates, 7 of S. marcescens 586 isolates and 4 of Acinetobacter spp. 474 isolates, respectively. Glycopeptide-resistant enterococci or S. aureus was not found.

[Analysis of small dense LDL in patients with type 2 diabetic mellitus by the modified Krauss method using an internal standard].

In patients with Type 2 diabetes mellitus (Type 2 DM), the relationship between the prevalence rate of small dense LDL (sdLDL) and parameters of lipid metabolism was analyzed using the method devised by modified Krauss method using apoferritin as an internal standard. The prevalence rate of sdLDL was 34% compared with it of normal subjects in this study. When the severity of Type 2 DM was classified into three groups of the HbA1c value, neither the sdLDL size nor its prevalence rate differed significantly depending upon the severity of the Type 2 DM. Also, when the prevalence rate of sdLDL was analyzed in relation to the severity of complications, i.e., of microangiopathy (retinopathy and nephropathy) or macroangiopathy (cerebral infarction), there was no significant difference in the prevalence rate of sdLDL depending on the severity of any of these complications. On the other hand, the prevalence rate of sdLDL was found to be correlated with the serum TG level. The serum level of TG-rich remnants (metabolites of TG) was also high in patients with sdLDL. It should take notice that the assessment of sdLDL should be used the authorized method for the evaluation. Thus it is concluded that the levels of sdLDL were important in evaluation of Type 2 DM. The prevalence rate of sdLDL did not correlate with the severity, nor the modalities for the complications of Type 2 DM.

Derivation of gender and age-specific reference intervals from fully normal Japanese individuals and the implications for health screening.

BACKGROUND: With nationwide standardization of laboratory tests among institutions for health screening in Japan, common reference intervals (RIs) were derived from records of 1,500,000 health check attendees. METHODS: Targets were 20 basic laboratory tests including body mass index (BMI) and systolic and diastolic blood pressures (SBP, DBP). Individuals fulfilling the following strict criteria were chosen: SBP<130, DBP<85mmHg, BMI<25kg/m(2), non-smoking, ethanol consumption<20g/day and under no mediation with no remarkable current/past illnesses. The latent abnormal values exclusion (LAVE) method was applied to ensure fully normal results. RIs were derived by parametric method using modified Box-Cox power transformation. RESULTS: Among all attendees, 23% fulfilled the criteria. Application of the LAVE method further reduced the dataset by 40%-50%. Age-related charts of test results differed greatly between genders in almost all tests. Comparison of derived RIs with clinical decision limits (CDLs) revealed that the upper limits of RIs differed from CDLs according to gender and age. CONCLUSIONS: Implementation of gender and age-specific RIs derived from individuals with fully normal health attributes will (1) enable appropriate interpretation of test results in health screening and (2) promote judicious application of CDLs for therapeutic intervention, taking into account gender, age and other health attributes.