Non-hybridization single nucleotide polymorphism detection and genotyping assay through direct discrimination of single base mutation by capillary electrophoretic separation of single-stranded DNA.

The significant demands for single nucleotide polymorphism detection and genotyping assays have grown. Most common assays are based on the recognition of the target sequence by the hybridization with its specific probe having the complementary sequence of the target. Herein, a simple, label-free, and economical non-hybridization assay was developed for single nucleotide polymorphism detection and genotyping, based on the direct discrimination of single base mutation by simple capillary electrophoresis separation for single-stranded DNA in an acidic electrophoretic buffer solution containing urea. Capillary electrophoresis separation of single-base sequential isomers of DNA was achieved due to charge differences resulting from the different protonation properties of the DNA bases. Single nucleotide polymorphism detection and genotyping were achieved by discriminating the electropherogram pattern change, that is, peak number in the electropherogram, obtained by the proposed method. The successful practical application of the proposed method was demonstrated through single nucleotide polymorphism detection and genotyping on a known gene region of 84-mer, in which guanine to adenine single-base mutation is commonly observed, using a human hair sample in combination with genomic DNA extraction, polymerase chain reaction amplification, DNA purification from polymerase chain reaction products, and capillary electrophoresis separation.

Separation of single-base sequential isomers of single-stranded DNA by capillary electrophoresis and its application in the discrimination of single-base DNA mutations.

Separation of single-base substitution sequential DNA isomers remains one of the most challenging tasks in DNA separation by capillary electrophoresis. We developed a simple, versatile capillary electrophoresis technique for the separation of single-base sequential isomers of DNA having the same chain length. This technique is based on charge differences resulting from the different protonation (acid dissociation) properties of the four DNA bases. A mixture of 13 single-base sequential isomers of 12-mer single-stranded DNA was separated by using an electrophoretic buffer solution containing 20 mM phosphoric acid (pH 2.0) and 8 M urea. We demonstrated that our method could separate all possible mutation patterns under identical experimental conditions. In addition, application of our method to the separation of the polymerase chain reaction product of a 68-mer gene fragment and its single-base isomers indicates that in combination with the appropriate genomic DNA extraction techniques, the method can detect single-base gene mutations.

Highly efficient near-infrared-emitting lanthanide(III) complexes formed by heterogeneous self-assembly of Ag(I), Ln(III), and thiacalix[4]arene-p-tetrasulfonate in aqueous solution (Ln(III) = Nd(III), Yb(III)).

Heterogeneous self-assembly of thiacalix[4]arene-p-tetrasulfonate (TCAS), Ag(I), and Ln(III) (= Nd(III), Yb(III)) in aqueous solutions conveniently afforded ternary complexes emitting Ln(III)-centered luminescence in the near-infrared (NIR) region. A solution-state study revealed that the Ag(I)-Nd(III)-TCAS system gave a complex Ag(I)(4)·Nd(III)·TCAS(2) in a wide pH range of 6-12. In contrast, the Ag(I)-Yb(III)-TCAS system gave Ag(I)(2)·Yb(III)(2)·TCAS(2) at a pH of around 6 and Ag(I)(2)·Yb(III)·TCAS(2) at a pH of approximately 9.5. The structures of the Yb(III) complexes were proposed based on comparison with known Ag(I)-Tb(III)-TCAS complexes that show the same self-assembly behavior. In Ag(I)(2)·Yb(III)(2)·TCAS(2), two TCAS ligands sandwiched a cyclic array of a Ag(I)-Ag(I)-Yb(III)-Yb(III) core. In Ag(I)(2)·Yb(III)·TCAS(2), Yb(III) was accommodated in an O(8) cube consisting of eight phenolate O(-) groups from two TCAS ligands linked by two S-Ag-S linkages. Crystallographic analysis of Ag(I)(4)·Nd(III)·TCAS(2) revealed that the structure was similar to Ag(I)(2)·Yb(III)·TCAS(2) but that it had four instead of two S-Ag-S linkages. The number of water molecules coordinating to Ln(III) (q) estimated on the basis of the luminescent lifetimes was as follows: Ag(I)(4)·Nd(III)·TCAS(2), 0; Ag(I)(2)·Yb(III)(2)·TCAS(2), 2.4; and Ag(I)(2)·Yb(III)·TCAS(2), 0. These findings were compatible with the solution-state structures. The luminescent quantum yield (Φ) for Ag(I)(4)·Nd(III)·TCAS(2) was 4.9 × 10(-4), which is the second largest value ever reported in H(2)O. These findings suggest that the O(8) cube is an ideal environment to circumvent deactivation via O-H oscillation of coordinating water. The Φ values for Ag(I)(2)·Yb(III)(2)·TCAS(2) and Ag(I)(2)·Yb(III)·TCAS(2) were found to be 3.8 × 10(-4) and 3.3 × 10(-3), respectively, reflecting the q value. Overall, these results indicate that the ternary systems have the potential for a noncovalent strategy via self-assembly of the multidentate ligand, Ln(III), and an auxiliary metal ion to obtain a highly efficient NIR-emissive Ln(III) complex that usually relies on elaborate covalent linkage of a chromophore and multidentate ligands to expel coordinating water.

Computerized surveillance system of tuberculosis in Japan: its evolution, achievement and challenges.

The nationwide computerized tuberculosis (TB) surveillance system was revised in 2007. It was developed to be user-friendly and to allow the evaluation of current TB problems and control issues in Japan. All public health centers in Japan (518 as of April 2007) have system terminals connected to a central computer, and the data entered at these terminals are sent to the online central computer excluding personal identification data. All the figures and tables in this paper were created using the annual report database which are compiled from this system. The revision in 2007 added many new functions to the system, such as a function for automatically sending data upon transfer. The monitoring information for assisting case management of TB patients by the DOTS was also enhanced. The algorithm for classifying treatment outcomes automatically based on data entered regarding cancellations from registration, bacteriological results and drug usage each month was revised. The proportion of "Failed" and "Defaulted" combined was 4.6% among new sputum smear positive pulmonary TB patients newly registered in 2009, while "Died" accounted for as high as 19.3%, due largely to a high percentage of the elderly. A new system for contact examination management is provided as a subsystem. Feedback of data analyses has been strengthened by various methods. This TB surveillance system is indispensable for implementing the evidence-based TB control program in Japan. An important role of the Research Institute of Tuberculosis is to support the planning and execution of TB control with provision of useful epidemiological information from the system.

[Present activity of BCG and LTBI treatment for infants in Japan].

I analyzed the BCG vaccination and treatment of latent tuberculosis infection (LTBI) for infants notified in 2008 in Japan. BCG was not recommended and treatment of LTBI was the main activity for prevention of TB in infants from birth to age 2 months. The majority of notified LTBI cases were detected by contact surveys. Out of the estimated number of TB infected (148), only 2 cases were notified based on the outcome of LTBI treatment for 89 infants. When the infants were 3-5 months old, both BCG vaccination and LTBI treatment were implemented. BCG coverage was 61.5-97.7%, and LTBI treatment for non-vaccinated individuals was applied for a larger number of infants (1.04 to 7.82 times as many) than the estimated number of infants infected with TB. The majority of infants were BCG vaccinated when they were 6-11 months old. Although LTBI treatment coverage was low, only 5 cases developed among those receiving BCG vaccination. During 1-2 year, BCG coverage was high and breakdown rates of BCG-vaccinated children were much lower than those of non-vaccinated children. This difference might be due to not only the preventive effect of BCG but also risk difference of TB infection between BCG-vaccinated and BCG non-vaccinated individuals. The number of notified LTBI treatment cases was lower than the estimated number of children infected with TB during 1-2 year. To prevent infant TB, reinforcement of contact surveys to identify LTBI for treatment, improvement of BCG coverage, and attention to BCG non-vaccinated infants older than 6 months might be effective.

[Recent case-finding activities of school children in Japan].

PURPOSE: The procedure for TB mass-screening of elementary and junior-high school students was revised in 2003 in Japan. An interview sheet with questions about the past history of TB, episodes of contact with TB patient, TB symptoms, and BCG vaccination history has been administered to all students to determine high-risk groups for TB infection or TB disease despite previous mass-screening with PPD testing for those in first grade. Our purpose in the present study was to evaluate the effects of revised case-finding activity for these school students. METHOD: We analyzed the case-finding activities for school students from 2003-2008 based on data from the national TB surveillance system and related ad-hoc surveys. RESULT: The total number of notified TB cases among these students was 295 out of which 156 were detected by contact survey, 110 by consultation at medical facilities with symptoms, 19 by mass-screening at schools, and the remaining 10 by other means. Although the contact investigation detected more than half of all cases, there were some more cases with a known source of infection among those who were detected in other modes; 28 cases were identified by consultation at medical facilities and 10 by school mass-screening. Case-finding activities by consultation at medical facilities detected 43 bacteriologically positive cases. There were 97 cases without a known source of infection. Most of the cases detected by school mass-examination had risk factors such as TB history in the family and/or a history of staying in TB-prevalent countries. The TB incidence in urban areas is higher than that in other areas for junior-high school students with known or unknown sources of infection. DISCUSSION: The effectiveness of the contact investigation could be enhanced by improving its way of implementation. Delay of diagnosis for infectious cases should be minimized as much as possible to prevent TB outbreaks among students. For that purpose, parents, school teachers, and school nurses should be aware to prevent a delay in seeking care for the patient. Because the efficiency of the current school mass-screening is quite low, revision of this program is necessary, taking into consideration the reinforcement of other case-finding activities. The difference between urban and other areas might be due to a higher infection risk in junior-high school students in urban areas who have higher social activity and may be exposed to higher TB infection risk due to casual contact.

[Trends in tuberculosis infection among foreigners in Japan according to work status].

PURPOSE: TB among foreigners is presently a serious issue in some developed countries and could become so in Japan. The purpose of this report is to assess the epidemiological situation of TB among foreigners in Japan. MATERIALS AND METHODS: The trend of TB reporting among foreigners in Japan was examined with regard to work status. RESULTS: The number of reported TB cases among employees and students in Japan increased between 1998 and 2008, but that among housekeepers was level throughout the same period. The increase among employees and students might be due to the increased numbers of foreign employees and students. In the case of housekeepers, the increase in the number of permanent residents did not lead to an increase in TB among these housekeepers. Estimates of TB reporting rates decreased during the study period, so the changes in reporting rates would not have caused the increase in TB cases. This downward trend may have been caused by an increase in longer-term residents and a decrease in TB incidence in home countries. Even though the TB reporting rate is decreasing, the rates in those countries are much higher than in Japan in the same work categories. DISCUSSION: To control the spread of TB, it is important to identify high-risk individuals. The Japanese TB control program should further strengthen mass health examination programs for foreign housekeepers and employees (especially temporary and daily employees), case-finding based on individuals' access to hospitals or clinics when suffering from TB symptoms, and flexible and periodic adjustment of TB control activities for foreigners according to future changes in the number and distribution of foreigners in Japan. Furthermore, improving the TB epidemiological situation in home countries might contribute to the downward trend of TB reporting rates among foreigners in Japan. Therefore, Japanese assistance in TB control activities in surrounding countries such as China, South Korea, and the Philippines might contribute to TB control activities for foreigners in Japan.

Ligand-substitution mode capillary electrophoretic reactor: extending capillary electrophoretic reactor toward measurement of slow dissociation kinetics with a half-life of hours.

A method employing capillary electrophoresis (CE) was developed to determine the rate constant of the very slow spontaneous dissociation of a complex species. The method uses a CE reactor (CER) to electrophoretically separate components from a complex zone and, thus, spontaneously dissociate a complex. The dissociation is accelerated by ligand substitution (LS) involving a competing ligand added to the electrophoretic buffer. The LS-CER method is validated using the dissociation of a Ti(IV)-catechin complex and EDTA as a competing ligand. There is good agreement between the spontaneous dissociation rate constant (k(d) = (1.64 +/- 0.63) x 10(-4) s(-1)) and the rate constant obtained by a conventional batchwise LS reaction (k(d) = (1.43 +/- 0.04) x 10(-4) s(-1)). Furthermore, the usefulness of the method is demonstrated using a Ti(IV)-tiron complex, for which k(d) = (0.51 +/- 0.43) x 10(-4) s(-1), corresponding to a half-life (t(1/2)) of 3.8 h. Notably, a single run of LS-CER for the Ti(IV) complex is completed within 40 min, implying that LS-CER requires a considerably shorter measurement time (roughly equal to t(1/2)) than conventional CER. LS-CER can be widely applied to determine the spontaneous dissociation rates of inorganic diagnostic and therapeutic reagents as well as of biomolecular complexes.

Expanding the scope of CE reactor to ssDNA-binding protein-ssDNA complexes as exemplified for a tool for direct measurement of dissociation kinetics of biomolecular complexes.

CE reactor (CER), which was developed as a tool for direct measurement of the dissociation kinetics of metal complexes, was successfully applied to the complexes of Escherichia coli ssDNA-binding protein (SSB) with ssDNA. The basic concept of CER is the application of CE separation process as a dissociation kinetic reactor for the complex, and the observation of the on-capillary dissociation reaction profile of the complex as the decrease of the peak height of the complex with increase of the migration time. The peak height of [SSB-ssDNA] decreases as the migration time increases since the degree of the decrease of [SSB-ssDNA] through the on-capillary dissociation reaction is proportional to the degree of the decrease of the peak height of [SSB-ssDNA]. The dissociation degree-time profiles for the complexes are quantitatively described by analyzing a set of electropherograms with different migration times. Dissociation rate constants of [SSB-ssDNA] consisting of 20-mer, 25-mer and 31-mer ssDNA were directly determined to be 3.99x10(-4), 4.82x10(-4) and 1.50x10(-3)/s, respectively. CER is a concise and effective tool for dissociation kinetic analysis of biomolecular complexes.

Separation of oligonucleotides with single-base mutation by capillary electrophoresis using specific interaction of metal ion with nucleotide.

A unique tactic for the separation of single-base sequential isomers of oligomeric single-stranded DNA by a CE separation system employing the specific interaction of metal ion with nucleotide was demonstrated, enabling the separation of the mixture of a 12-mer oligonucleotide and its single-base mutants, as well as their positional isomers.

Capillary electrophoretic separation of humic substances using hydroxyethyl cellulose as a buffer additive and its application to characterization of humic substances in a river water sample.

We have developed a concise tool for the investigation of the transition of humic substances in environmental water. The separation of water-soluble humic substances was achieved rapidly and effectively by capillary electrophoresis using a polyacrylamide-coated capillary and a phosphate electrophoretic buffer solution (pH 7.0) containing hydroxyethyl cellulose. The separation mechanism was assessed using the ultrafiltration technique. The effect of the complexation of humic substances with metal ions was studied by using the proposed method. When Fe(III) ions or EDTA was added to the sample solution of fulvic acid, a distinct change in the electropherogram pattern based on the conformational change of fulvic acid was observed. The successful application of the proposed method to the characterization of humic substances in a river water sample was also demonstrated.

[Comparison of TB incidence of young and middle age groups between urban/suburban prefectures and other prefectures].

PURPOSE: Comparison of TB incidence in young and middle age group between urban/surrounding area and other area. MATERIAL & METHODS: TB incidence in young and middle age group by sex, nationality, labor status and prefecture is calculated with data of National Population Survey and TB surveillance in 2005. Risk factors of TB infection/break down such as public transportation, poverty, population density, diabetes, malignancy, HIV infection, foreigners are compared between urban/surrounding area and other area. RESULTS: TB incidence of employees, jobless and housekeepers in urban/surrounding prefectures is higher than other prefectures, but TB incidence of housekeepers is low and difference is small. Utilization frequency of public transportation is correlated with prefectural level of TB incidence of male and female employees. There is no correlation with prevalence of other risk factors. DISCUSSION: Public transportation and working in urban area is indicated as causes of difference of TB infection risk. Reinforcement of TB control program of working places in urban area is important and if necessary contact survey of TB outbreak at public transportation should be considered. Further analysis using more precise indicators is necessary to elucidate for influence of poverty.

[Influence of occupation of the index cases in recent outbreaks of tuberculosis].

UNLABELLED: PURPOSE, MATERIALS AND METHODS: In order to elucidate risk factors of TB outbreaks, 223 outbreaks reported between 2000 and 2006 in Japan were analyzed according to sex, occupation and outbreaking place of the index case. RESULTS: In male, the most frequent occupation was permanent workers who infected contacts mainly at workplaces, followed by unemployed people at general hospital or nursing home for the aged. The third was students who infected contacts mostly at school, followed by customer relations workers at companies or amusement places. In female, the most frequent occupation was students who infected contacts at school. Second most frequent was unemployed who infected contacts at general hospitals and nursing homes for the aged, followed by nurses at general hospitals or mental hospitals. Frequent outbreak places were workplaces, schools, general hospitals, amusement places and mental hospitals in male, and general hospitals and schools in female. The risk of becoming the source of TB outbreak was high in teachers/doctors, students, unemployed as well as customer relations workers in male, and teachers/doctors, students as well as nurses in female. DISCUSSION: The most important point to prevent outbreaks of TB is to find out TB suspects with relevant symptoms for tuberculosis as early as possible to minimize patient's delay, especially from people with high risk of TB infection to others such as teachers/doctors, students and medical nurses. As most frequent job and place were general permanent workers at workplaces, TB control activities to minimize patient's delay at general workplace is also important.

[A case of multiple-drug-resistant pulmonary tuberculosis in a hemodialysis hospital].

A 42-year-old male with chronic renal failure was diagnosed as multiple-drug-resistant pulmonary tuberculosis, 9 months after the onset of hemodialysis. During the period before the diagnosis of tuberculosis, he visited regularly the hemodialysis hospital, accordingly many patients with hemodialysis and the hospital staffs had close and long contact with the patient. Our health center planned and conducted contact examinations among them, with QFT-TB test and chest X-ray. Petients with hemodialysis are regarded as immunocompromised hosts, one of the high risk groups for infections. The result of QFT-TB test of patients were negative, however, we have to cosider the possibility of false-negative. So we followed up by monthly chest X-ray examination all hemodialysis patients for 2 years, and finally, it was found that secondary infection of multiple-drug-resistant tuberculosis among immunocompromised hosts did not occur.

[Generation-to-generation TB transmission in Aichi, Central Japan -- an epidemiological study of 701 TB patients in 290 clusters].

OBJECTIVES: To elucidate characteristics in generation-to-generation TB transmission from smear positive pulmonary TB patients. SUBJECTS AND METHODS: The subjects of this retrospective study were 10,088 TB patients registered in Aichi prefecture between 1989 and 2003. Smear-positive pulmonary TB was found in 3,332. All registration files were reviewed to identify epidemiological links of patients. When linked patients with an interval of the dates of registration of less than 10 years were found, the first case was considered as the index case, and the other patients were regarded as secondary patients. All patients were subdivided by every 10 years of age as a generation. All patients were also grouped as young generations aged 0 to 29, middle-aged generations aged 30 to 59, and elder generations aged 60 to 99. A young generations rate (YGR) for a generation of index cases was defined as following; YGR=NY/NA, where NA: Number of whole TB secondary patients transmitted from all index cases belonging to generation A, and NY: Number of secondary patients aged 0-29 in NA. A same generation rate (SGR) for a generation of index cases was defined as following; SGR=NS/NA, where NA: Number of whole TB secondary patients transmitted from all index cases belonging to generation A, and NS: Number of secondary patients belonging to the same generation A in NA. RESULTS: A total of 290 clusters were found with 290 index cases and 411 secondary cases. Of the 290 index cases, the biggest number of patients was 58 for the forties, followed by 43 patients for the twenties and the fifties. Of the 411 secondary patients, the biggest number of patients was 91 for the twenties, followed by 58 patients for the thirties, and 158 patients or 38.4% belonged to the young generations. High YGRs were found in the young and middle-aged generations ranging from 37.5% to 75.0%, while YGRs were low in the elder generations ranging from 15.9% to 26.7%. The difference was significant between the fifties and the sixties (p<0.01). The YGRs were calculated 57.1% for the young generations, 43.5% for the middle-aged generations, and 18.9% for the elder generation. The differences were significant between the young and the middle-aged (p<0.05), as well as between the middle-aged and the elder (p<0.001). The biggest SGR was calculated 42.2% for the twenties. Low SGRs were 18.2% for the forties and 17.5% for the fifties. The differences were significant between the twenties and the forties (p<0.01), as well as between the twenties and the fifties (p<0.01). The secondary patients aged 0 to 29 were counted 158: 123 were transmitted through intra-familial infection and 35 were through extra-familial infection. The major index case generation was the middle-aged generations with 82 or 66.7% secondary patients in the case of intra-familial infection, while it was the young generations with 20 or 57.1% patients in the case of extra-familial infection. CONCLUSION: These findings suggest that the major sources of TB transmission to young people are smear-positive pulmonary TB patients aged 0 to 59.

Determination of boron in water samples at nanograms per cubic decimeter levels by reversed-phase partition high-performance liquid chromatography with precolumn complexation reaction using salicylaldehyde and 1-amino-8-naphthol-3,6-disulfonate.

A new method for the highly sensitive and selective determination of boron at nanograms per cubic decimeter levels has been developed based on the derivatization reaction of boron using salicylaldehyde and 1-amino-8-naphtol-3,6-disulfonate with reversed-phase partition high-performance liquid chromatography. A detection limit as low as 2.0 nmol/dm(3) (22 ng/dm(3)) was achieved without any preconcentration. No significant interference was observed in the determination of 16 micromol/dm(3) of boron with the addition of nine metal ions (Al(III), Cu(II), Co(II), Fe(II), Fe(III), Ni(II), Mn(II), V(V), Zn(II)) at concentrations 100 times greater than that of boron without any masking procedure. The proposed method was successfully applied to the determination of boron in river water, tap water, doubly distilled water, and highly purified water.

[TB epidemiology in an elderly group in Japan].

PURPOSE: To analyze the epidemiological situation of TB among the elderly in Japan. METHODS: By using the data of TB surveillance, national survey statistics and national basic survey of life. RESULTS: TB incidence in both sexes increased with age in all survey years. TB incidence among the jobless was higher than among employees in both sexes. TB incidence among female housekeepers was consistently lower than among other groups. TB incidence among both the employed and unemployed has declined rapidly in the last 15 years. The main mode of TB detection was the detection at clinics/hospitals irrespective of presence or absence of TB symptoms. The death rate was higher in cases with immunosuppressive therapy and cases with malignancy. CONCLUSION: Causes of higher TB incidence in the elderly unemployed population should be analyzed. Better detection of TB by improving passive case-finding for TB symptomatic and high-risk groups at clinics/hospitals will be recommended. To reduce the death rate, early diagnosis of TB with immunosuppressive therapy could be strengthened, however its impact might be limited.

[Relapse rate of pulmonary tuberculosis within two years following completion of twice weekly intermittent chemotherapy].

OBJECTIVE: To evaluate the relapse rate after treatment of a twice weekly intermittent chemotherapy during continuation phase in the patients with pulmonary tuberculosis in two years period after completion of chemotherapy. METHODS: The patients with drug susceptible pulmonary tuberculosis treated with 2HRZE/4H2R2 under the supervision by the pharmacists or the patients treated with 2HRZE/4HR by self-administration were followed-up for two years after completion of chemotherapy. RESULTS: A total number of 135 pulmonary tuberculosis patients were treated with 2HRZE/4H2R2, 3 of 135 discontinued this intermittent treatment, and 11 of 135 completed chemotherapy was excluded from the relapse analysis, 105 out of the remaining 121 were followed-up for more than 6 months. On the other hand 240 patients were treated with 2HRZE/4HR, 37 out of 240 were excluded from the analysis, 147 of the remaining 203 were followed-up for more than 6 months. The relapse rate of this intermittent chemotherapy 1.89/100 person-year was similar to the relapse rate 1.86/100 person-year among 147 treated with daily regimen by self-administration. This difference was not statistically significant (z = 0.36, P = 0.14). CONCLUSION: As regards relapse rate, this 2HRZE/4H2R2 regimen is effective and useful for the expansion of DOT, and it should be expanded nationally in Japan.

[Tuberculosis control in Shinjuku Ward, Tokyo--promoting the DOTS program and its outcome].

OBJECTIVES: The objectives were to report how to promote tuberculosis (TB) control including DOTS (Directly Observed Treatment, Short-course) programs, and to evaluate the results of TB control programs in Shinjuku Ward (Shinjuku-ku). SETTING AND CHARACTERISTICS: Inhabitants and TB patients in Shinjuku Ward. Shinjuku Ward is located in the center of metropolitan Tokyo and has typical urban TB problems, such as high incidence rate and TB among foreigners and the homeless. The TB incidence rates in Shinjuku Ward decreased from 83.9 per 100,000 population in 1999 to 42.5 per 100,000 population in 2006, however, the rates were still two times higher than the national average. Therefore, one of the important TB programs in Shinjuku has been to actively detect cases among high-risk groups such as foreigners and the homeless. METHODS: We observed the trend of case detection rates by health examination with chest X-ray among different high-risk groups, and compared the treatment outcomes before and after DOTS program execution. We also reviewed the changes of re-treatment rates and drug resistance rates. RESULTS: The case detection rates of TB by health examinations of foreign students at Japanese language schools decreased from 0.49% in 1996 to 0.13% in 2006 (p = 0.021). Although the case detection rates decreased, they were still about 26 times higher than those of Japanese students. While, the case detection rates among the homeless remained high with 4.7%, 3.3%, 4.5% and 3.6% in 1999-2002, respectively, since 2003, however, they had decreased and no TB cases were detected in 2005-2006. The DOTS program for homeless TB patients has been carried out since 2000 and that for the foreigners since 2003. The rates of defaulting during treatment before DOTS were very high among both homeless patients (21.4%) and foreigners (29.8%) in 1998-1999. However, after the introduction of DOTS program, those rates declined to 10.4% (p = 0.014) among the homeless and 7.8% (p = 0.002) among foreigners in 2002-2004. The proportion of newly notified patients with previous TB treatment and those with multi-drug resistant TB (MDR-TB) have also decreased after the introduction of DOTS programs. From 2000-2002 to 2003-2006, the re-treatment rates decreased from 19.4% to 10.0% (p < 0.001) and MDR-TB rates decreased from 1.6% to 0.2% (p = 0.042), respectively. DISCUSSION: The key points of TB control in Shinjuku Ward are to detect TB cases early especially among the high-risk groups, and to assist all TB patients to complete their treatment. In order to expand this strategy, besides promoting active case findings among high-risk groups, we have developed many types of DOTS programs, considering each patient's lifestyle and cooperating with school teachers at schools, pharmacists at pharmacies, home-care specialists at homes or facilities for the elderly, and so on. Among others, a major premise for the homeless and some other socially disadvantaged patients was to guarantee the provision of medicine and living by introducing social welfare services, before starting DOTS programs. This approach might have helped to reduce the defaulting rate, relapse rate and MDR-TB rate.

Safe and rapid development of capillary electrophoresis for ultratrace uranyl ions in radioactive samples by way of fluorescent probe selection for actinide ions from a chemical library.

After the serious nuclear accident at the Fukushima Daiichi Nuclear Power Plant caused by the Great East Japan Earthquake in 2011, the development of feasible, safe, and highly sensitive analytical methods (in terms of low levels of radiation exposure and radioactive waste generation) for radioactive samples, especially actinide (An) ions, represents an important challenge. Here we propose a methodology for selecting appropriate emissive probes for An ions with very low consumption and emission of radioactivity by capillary electrophoresis-laser-induced fluorescence detection (CE-LIF), using a small chemical library of probes with eight different chelating moieties. It was found that the emissive probe L1, which possesses the tetradentate chelating moiety 1,10-phenanthroline-2,9-dicarboxylic acid (PDA), was suitable for detecting uranyl ions. The detection limit for the uranyl-L1 complex using CE-LIF combined with dynamic ternary complexation and on-capillary concentration techniques was determined to be 2.9 × 10-12 M (0.7 ppt). No interference from the large excess of matrix metal ions was observed. This method was successfully applied to real radioactive liquid samples collected from nuclear facilities, including the Fukushima Daiichi Nuclear Power Plant. This strategy not only permitted the development of a safe and rapid analytical method but also provided insight into the coordination chemistry of An ion complexes. Specifically, the PDA structure provided substantial kinetic inertness to its uranyl complex; the formation of a ternary complex between uranyl-L1 and carbonate was revealed; and unusual interactions were observed between the π-electron systems of uranyl and the phenanthroline ring, which stabilized the uranyl-PDA interaction.