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Sample records for afb smear-positive sputum

  1. Cost-effectiveness of chemotherapy for sputum smear-positive pulmonary tuberculosis in Malawi, Mozambique and Tanzania.

    PubMed

    de Jonghe, E; Murray, C J; Chum, H J; Nyangulu, D S; Salomao, A; Styblo, K

    1994-01-01

    The cost-effectiveness of chemotherapy for pulmonary sputum smear-positive tuberculosis was examined in the national tuberculosis control programmes of Malawi, Mozambique and Tanzania. In these three programmes, routine cure rates have exceeded 80 per cent. Average, average incremental and marginal unit costs for standard, short-course and retreatment regimens with and without hospitalization have been measured. The average incremental cost per year of life saved through chemotherapy ranged from US $0.90-3.10. In all conditions, short-course chemotherapy is preferable to standard 12-month chemotherapy. When hospitalization during the intensive phase of chemotherapy increases the cure rate by 10-15 percentage points, it can be relatively cost-effective. Analysing the cost-effectiveness of short-course and standard chemotherapy, where the depth of the margin of benefit is different, illustrates some of the dangers of simplistic use of cost-effectiveness ratios.

  2. Rate of recovery of Mycobacterium tuberculosis from frozen acid-fast-bacillus smear-positive sputum samples subjected to long-term storage in Northwest Ethiopia.

    PubMed

    Tessema, Belay; Beer, Joerg; Emmrich, Frank; Sack, Ulrich; Rodloff, Arne C

    2011-07-01

    Tuberculosis is a major public health problem in Ethiopia. The diagnosis and treatment of drug-resistant tuberculosis remain a challenge in the country. This study aimed to assess whether single morning sputum samples could be stored at -20 °C for extended periods of time at remote settings and then transported and successfully cultured for Mycobacterium tuberculosis. Single morning sputum samples were collected from all smear-positive tuberculosis patients diagnosed at Gondar Hospital, Gondar Health Center, Metemma Hospital, Bahir Dar Hospital, and Debre Markos Hospital in Northwest Ethiopia between March and July 2009. Specimens were stored at the study sites and sent to the mycobacteriology laboratory at the University Hospital, Leipzig, Germany, where specimens were processed and inoculated into the BacT/Alert 3D system and Lowenstein-Jensen and Gottsacker media. Ice packs were added in the package of the specimens during transport. A total of 319 patients were enrolled in this study. The median specimen storage time was 132 days (range, 16 to 180 days). Of all specimens, 283 (88.7%) were culture positive by any of the three culturing systems. M. tuberculosis isolates from four contaminated specimens in all culturing systems were successfully isolated on Middlebrook 7H10 agar; thereby, the recovery rate increased to 287 (90.0%). The length of time of sputum storage had no significant effect on the rate of recovery of M. tuberculosis in all culturing systems. In conclusion, single morning sputum specimens collected at remote settings stored at -20 °C for long periods of time without the addition of preservatives can yield a high recovery rate. These findings suggest a simple and cost-effective alternative method of sputum storage for epidemiological and drug resistance studies in low-resource countries. PMID:21562105

  3. Rate of Recovery of Mycobacterium tuberculosis from Frozen Acid-Fast-Bacillus Smear-Positive Sputum Samples Subjected to Long-Term Storage in Northwest Ethiopia ▿

    PubMed Central

    Tessema, Belay; Beer, Joerg; Emmrich, Frank; Sack, Ulrich; Rodloff, Arne C.

    2011-01-01

    Tuberculosis is a major public health problem in Ethiopia. The diagnosis and treatment of drug-resistant tuberculosis remain a challenge in the country. This study aimed to assess whether single morning sputum samples could be stored at −20°C for extended periods of time at remote settings and then transported and successfully cultured for Mycobacterium tuberculosis. Single morning sputum samples were collected from all smear-positive tuberculosis patients diagnosed at Gondar Hospital, Gondar Health Center, Metemma Hospital, Bahir Dar Hospital, and Debre Markos Hospital in Northwest Ethiopia between March and July 2009. Specimens were stored at the study sites and sent to the mycobacteriology laboratory at the University Hospital, Leipzig, Germany, where specimens were processed and inoculated into the BacT/Alert 3D system and Lowenstein-Jensen and Gottsacker media. Ice packs were added in the package of the specimens during transport. A total of 319 patients were enrolled in this study. The median specimen storage time was 132 days (range, 16 to 180 days). Of all specimens, 283 (88.7%) were culture positive by any of the three culturing systems. M. tuberculosis isolates from four contaminated specimens in all culturing systems were successfully isolated on Middlebrook 7H10 agar; thereby, the recovery rate increased to 287 (90.0%). The length of time of sputum storage had no significant effect on the rate of recovery of M. tuberculosis in all culturing systems. In conclusion, single morning sputum specimens collected at remote settings stored at −20°C for long periods of time without the addition of preservatives can yield a high recovery rate. These findings suggest a simple and cost-effective alternative method of sputum storage for epidemiological and drug resistance studies in low-resource countries. PMID:21562105

  4. Primary Multidrug Resistant Tuberculosis and Utility of Line Probe Assay for Its Detection in Smear-Positive Sputum Samples in a Tertiary Care Hospital in South India

    PubMed Central

    Yacoob, Fahmiya Leena; Philomina Jose, Beena; Karunakaran Lelitha, Sarada Devi; Sreenivasan, Sreelatha

    2016-01-01

    In a high tuberculosis burdened country like India, rapid, cost-effective, and reliable diagnostic tools for tuberculosis are an urgent need of the hour to prevent inappropriate treatment strategies and further spread of resistance. This study aimed to estimate the proportion of new smear-positive tuberculosis cases with primary resistance to rifampicin and/or isoniazid as well as identify the common mutations associated with it. Sputum of 200 newly diagnosed smear-positive cases of 1+ score and above was directly subjected to Line Probe Assay using the GenoType MTBDRplus assay kit. All samples were inoculated onto solid media and 61 samples were inoculated in automated liquid culture also. The Line Probe Assay gave hundred percent interpretable results with 2.5% of the study population showing resistant pattern. Only 1% of the cases were primary multidrug resistant tuberculosis and 1.5% showed isoniazid monoresistance. S531L and C15T were the most common genetic mutations seen for rifampicin and isoniazid resistance, respectively. 40% had absent rpoB wild type 8 band indicating probable silent mutation after clinical correlation. The average turnaround time for Line Probe Assay was far less (3.8 days) as compared to solid and liquid cultures (35.6 days and 13.5 days, resp.). PMID:27099794

  5. Correlation between tuberculin skin test and IGRAs with risk factors for the spread of infection in close contacts with sputum smear positive in pulmonary tuberculosis

    PubMed Central

    2014-01-01

    Background The aim of the study was to assess the correlation between the tuberculin skin test (TST) and in vitro interferon-gamma released assays (IGRAs) with risk factors for the spread of infection in smear positive pulmonary tuberculosis (TB) contacts. Methods We recruited prospective contacts with smear positive pulmonary TB cases. We looked at human immunodeficiency virus (HIV) infection and other conditions of immunosuppression, presence of BCG vaccination and the degree of exposure to the index case. Patients underwent the TST, chest radiography, sputum analysis when necessary, and IGRA assays (QFN-G-IT and T-SPOT.TB). Presence of cough, diagnostic delay (days between first symptoms and TB diagnostic), contact conditions: room size (square meters) and index of overcrowding (square meters per person) were investigated in the index case. Results 156 contacts (119 adults, 37 children) of 66 TB patients were enrolled, 2.4 (1-14) contacts per TB case. The positivity of the TST did not correlate with the risk factors studied: presence of cough (p = 0.929); delayed diagnosis (p = 0.244); room size (p = 0.462); overcrowding (p = 0.800). Both QFN-G-IT and T-SPOT.TB, showed significant association with cough (p = 0.001, and p = 0.007) and room size (p = 0.020, and p = 0.023), respectively. Conclusions Both IGRA associated better than TST with certain host-related risk factors involved in the transmission of disease, such as the presence of cough. PMID:24885850

  6. Comparative proteomic analysis of serum diagnosis patterns of sputum smear-positive pulmonary tuberculosis based on magnetic bead separation and mass spectrometry analysis.

    PubMed

    Liu, Jiyan; Jiang, Tingting; Jiang, Feng; Xu, Dandan; Wei, Liliang; Wang, Chong; Chen, Zhongliang; Zhang, Xing; Li, Jicheng

    2015-01-01

    A major challenge in pulmonary tuberculosis (TB) control is early and accurate diagnosis of sputum smear negative pulmonary TB (SSN-PTB). The patients with SSN-PTB have to wait for a longer period of time before receiving proper treatment than sputum smear positive pulmonary TB (SSP-PTB) patients due to delay in diagnosis. The purpose of this study is to discover potential serum protein biomarkers for SSN-PTB. Surface-enhanced laser desorption/ionization-time of flight mass spectrometry (SELDI-TOF MS) combined with weak cation exchange (WCX) magnetic beads was used to screen serum samples from SSN-PTB patients (N = 66), SSP-PTB patients (N = 49), and healthy volunteers (N = 80). The serum protein profiles were analyzed with Biomarker Wizard system. A classification model was established using Biomarker Pattern Software (BPS). Fifty-eight protein peaks were identified to exhibit significant differences between SSN-PTB, SSP-PTB and healthy control groups (P < 0.05), among which 6 peaks were found to be down-regulated, while 10 peaks were up-regulated gradually in the healthy control, SSN-PTB, and SSP-PTB groups. Twenty-three discriminating m/z peaks were detected between SSN-PTB patients and healthy controls (P < 0.01, Fold ≥ 1.5). The classification tree combined with three protein peaks (2747.0, 4480.0, and 9410.1 Da) could distinguish SSN-PTB patients from healthy controls with a sensitivity of 83.33% and a specificity of 82.50%. Early diagnosis of SSN-PTB disease is critical in order to reduce morbidity and mortality associated with TB. The study will help to clarify the role of differential proteins in the pathogenesis of TB.

  7. Randomized dose-ranging study of the 14-day early bactericidal activity of bedaquiline (TMC207) in patients with sputum microscopy smear-positive pulmonary tuberculosis.

    PubMed

    Diacon, Andreas H; Dawson, Rodney; Von Groote-Bidlingmaier, Florian; Symons, Gregory; Venter, Amour; Donald, Peter R; Conradie, Almari; Erondu, Ngozi; Ginsberg, Ann M; Egizi, Erica; Winter, Helen; Becker, Piet; Mendel, Carl M

    2013-05-01

    Bedaquiline is a new antituberculosis agent targeting ATP synthase. This randomized, double-blinded study enrolling 68 sputum smear-positive pulmonary tuberculosis patients evaluated the 14-day early bactericidal activity of daily doses of 100 mg, 200 mg, 300 mg, and 400 mg bedaquiline, preceded by loading doses of 200 mg, 400 mg, 500 mg, and 700 mg, respectively, on the first treatment day and 100 mg, 300 mg, 400 mg, and 500 mg on the second treatment day. All groups showed activity with a mean (standard deviation) daily fall in log10 CFU over 14 days of 0.040 (0.068), 0.056 (0.051), 0.077 (0.064), and 0.104 (0.077) in the 100-mg, 200-mg, 300-mg, and 400-mg groups, respectively. The linear trend for dose was significant (P = 0.001), and activity in the 400-mg dose group was greater than that in the 100-mg group (P = 0.014). All of the bedaquiline groups showed significant bactericidal activity that was continued to the end of the 14-day evaluation period. The finding of a linear trend for dose suggests that the highest dose compatible with safety considerations should be taken forward to longer-term clinical studies. PMID:23459487

  8. Randomized Dose-Ranging Study of the 14-Day Early Bactericidal Activity of Bedaquiline (TMC207) in Patients with Sputum Microscopy Smear-Positive Pulmonary Tuberculosis

    PubMed Central

    Dawson, Rodney; Von Groote-Bidlingmaier, Florian; Symons, Gregory; Venter, Amour; Donald, Peter R.; Conradie, Almari; Erondu, Ngozi; Ginsberg, Ann M.; Egizi, Erica; Winter, Helen; Becker, Piet; Mendel, Carl M.

    2013-01-01

    Bedaquiline is a new antituberculosis agent targeting ATP synthase. This randomized, double-blinded study enrolling 68 sputum smear-positive pulmonary tuberculosis patients evaluated the 14-day early bactericidal activity of daily doses of 100 mg, 200 mg, 300 mg, and 400 mg bedaquiline, preceded by loading doses of 200 mg, 400 mg, 500 mg, and 700 mg, respectively, on the first treatment day and 100 mg, 300 mg, 400 mg, and 500 mg on the second treatment day. All groups showed activity with a mean (standard deviation) daily fall in log10 CFU over 14 days of 0.040 (0.068), 0.056 (0.051), 0.077 (0.064), and 0.104 (0.077) in the 100-mg, 200-mg, 300-mg, and 400-mg groups, respectively. The linear trend for dose was significant (P = 0.001), and activity in the 400-mg dose group was greater than that in the 100-mg group (P = 0.014). All of the bedaquiline groups showed significant bactericidal activity that was continued to the end of the 14-day evaluation period. The finding of a linear trend for dose suggests that the highest dose compatible with safety considerations should be taken forward to longer-term clinical studies. PMID:23459487

  9. Randomized dose-ranging study of the 14-day early bactericidal activity of bedaquiline (TMC207) in patients with sputum microscopy smear-positive pulmonary tuberculosis.

    PubMed

    Diacon, Andreas H; Dawson, Rodney; Von Groote-Bidlingmaier, Florian; Symons, Gregory; Venter, Amour; Donald, Peter R; Conradie, Almari; Erondu, Ngozi; Ginsberg, Ann M; Egizi, Erica; Winter, Helen; Becker, Piet; Mendel, Carl M

    2013-05-01

    Bedaquiline is a new antituberculosis agent targeting ATP synthase. This randomized, double-blinded study enrolling 68 sputum smear-positive pulmonary tuberculosis patients evaluated the 14-day early bactericidal activity of daily doses of 100 mg, 200 mg, 300 mg, and 400 mg bedaquiline, preceded by loading doses of 200 mg, 400 mg, 500 mg, and 700 mg, respectively, on the first treatment day and 100 mg, 300 mg, 400 mg, and 500 mg on the second treatment day. All groups showed activity with a mean (standard deviation) daily fall in log10 CFU over 14 days of 0.040 (0.068), 0.056 (0.051), 0.077 (0.064), and 0.104 (0.077) in the 100-mg, 200-mg, 300-mg, and 400-mg groups, respectively. The linear trend for dose was significant (P = 0.001), and activity in the 400-mg dose group was greater than that in the 100-mg group (P = 0.014). All of the bedaquiline groups showed significant bactericidal activity that was continued to the end of the 14-day evaluation period. The finding of a linear trend for dose suggests that the highest dose compatible with safety considerations should be taken forward to longer-term clinical studies.

  10. A Novel Reading Scheme for Assessing the Extent of Radiographic Abnormalities and Its Association with Disease Severity in Sputum Smear-Positive Tuberculosis: An Observational Study in Hyderabad/India

    PubMed Central

    Goyal, Surabhi; Hussain, Abid; Klassert, Tilman E.; Driesch, Dominik; Tokaryeva, Viktoriya; Löschmann, Yvonne Yi-Na; Sumanlatha, Gadamm; Ahmed, Niyaz; Valluri, Vijayalakshmi; Schumann, Ralf R.; Lala, Birgit; Slevogt, Hortense

    2015-01-01

    Background Existing reading schemes for chest X-ray (CXR) used to grade the extent of disease severity at diagnosis in patients with pulmonary tuberculosis (PTB) are often based on numerical scores that summate specific radiographic features. However, since PTB is known to exhibit a wide heterogeneity in pathology, certain features might be differentially associated with clinical parameters of disease severity. Objective We aimed to grade disease severity in PTB patients at diagnosis and after completion of DOTS treatment by developing a reading scheme based on five different radiographic manifestations and analyze their association with the clinical parameters of systemic involvement and infectivity. Methods 141 HIV-negative adults with newly diagnosed sputum smear-positive PTB were enrolled in a prospective observational study in Hyderabad, India. The presence and extent on CXRs of five radiographic manifestations, i.e., lung involvement, alveolar infiltration, cavitation, lymphadenopathy and pleural effusion, were classified using the new reading scheme by using a four-quadrant approach. We evaluated the inter-reader reliability of each manifestation, and its association with BMI and sputum smear positivity at diagnosis. The presence and extent of these radiographic manifestations were further compared with CXRs on completion of DOTS treatment. Results At diagnosis, an average lung area of 51.7% +/- 23.3% was affected by radiographic abnormalities. 94% of the patients had alveolar infiltrates, with 89.4% located in the upper quadrants, suggesting post primary PTB and in 34.8% of patients cavities were found. We further showed that the extent of affected lung area was a negative predictor of BMI (β value -0.035, p 0.019). No significant association of BMI with any of the other CXR features was found. The extent of alveolar infiltrates, along with the presence of cavitation, were strongly associated with sputum smear positivity. The microbiological cure rate in

  11. Decentralization of Acid Fast Bacilli(AFB) External Quality Assurance Using Blind Rechecking for Sputum Smear Microscopy in Ethiopia

    PubMed Central

    Melese, Muluken; Jerene, Degu; Alem, Genetu; Seid, Jemal; Belachew, Feleke; Kassie, Yewulsew; Habte, Dereje; Negash, Solomon; Ayana, Gonfa; Girma, Belaineh; Haile, Yared K.; Hiruy, Nebiyu; Suarez, Pedro G.

    2016-01-01

    Introduction Ethiopia achieved a rapid expansion of TB microscopic centers for acid fast bacilli (AFB). However, external quality assurance (EQA) services were, until recently, limited to few regional and sub-regional laboratories. In this paper, we describe the decentralization experience and the result of EQA using random blinded rechecking. Materials and Methods The routine EQA quarterly report was compiled and analyzed. A positive result by the microscopic center while the EQA center reported negative result is categorized as false positive (FP). A negative result by the microscopic center while the EQA center reported positive is considered false negative (FN). The reading of EQA centers was considered a gold standard to compute the sensitivity, specificity, positive predictive (PPV) and negative predictive values (NPV) of the readings of microscopic centers. Results We decentralized sputum smear AFB EQA from 4 Regional Laboratories (RRLs) to 82 EQA centers and enrolled 956 health facilities in EQA schemes. Enrollment of HFs in EQA was gradual because it required training and mentoring laboratory professionals, institutionalizing internal QA measures, equipping all HFs to perform diagnosis, and establishing more EQA centers. From 2012 to 2014 (Phase I), the FP rate declined from 0.6% to 0.2% and FN fell from as high as 7.6% to 1.6% in supported health facilities (HFs). In HFs that joined in Phase II, FN rates ranged from 5.6 to 7.3%. The proportion of HFs without errors has increased from 77.9% to 90.5% in Phase I HFs and from 82.9% to 86.9% in Phase II HFs. Overall sensitivity and specificity were 95.0% and 99.7%, respectively. PPV and NPV were 93.3% and 99.7%, respectively. Conclusion Decentralizing blinded rechecking of sputum smear microscopy is feasible in low-income settings. While a comprehensive laboratory improvement strategy enhanced the quality of microscopy, laboratory professionals’ capacity in slide reading and smear quality requires continued

  12. Clinical significance of smear positivity for acid-fast bacilli after ≥5 months of treatment in patients with drug-susceptible pulmonary tuberculosis

    PubMed Central

    Kang, Hyung Koo; Jeong, Byeong-Ho; Lee, Hyun; Park, Hye Yun; Jeon, Kyeongman; Huh, Hee Jae; Ki, Chang-Seok; Lee, Nam Yong; Koh, Won-Jung

    2016-01-01

    Abstract Patients with pulmonary tuberculosis (TB) with acid-fast bacilli (AFB)-positive sputum smear at 5 months or later during treatment are considered to be cases of treatment failure according to World Health Organization guidelines. This study evaluated the proportion, clinical characteristics, and significance of positive sputum smears after ≥5 months of standard treatment in patients with drug-susceptible pulmonary TB. This was a retrospective cohort study of 1611 patients with culture-confirmed drug-susceptible pulmonary TB who received standard anti-TB treatment from January 2009 to February 2014. Forty-one patients (2.5%) who were smear-positive after ≥5 months of treatment and 123 age- and sex-matched control patients were evaluated. Among the 41 smear-positive patients, culture of the sputum specimens yielded Mycobacterium tuberculosis (MTB) in 1 patient (2.4%), nontuberculous mycobacteria (NTM) in 7 (17.1%), and no growth in the remaining 33 patients (80.5%). Treatment was successfully completed in 40 patients (97.6%) with prolongation of the continuation phase regimens without change to second-line anti-TB treatment. In patients with smear positivity after ≥5 months of treatment compared with controls, cavitation on chest radiographs (53.7% vs. 25.2%, P = 0.001), bilateral involvement (51.2% vs. 30.1%, P = 0.01) and combined pleural effusion (26.8% vs. 10.6%, P = 0.01) were found more frequently at the time of treatment initiation, and paradoxical response occurred more commonly (19.5% vs. 3.3%, P = 0.002) during treatment. Smear-positive sputum after ≥5 months of standard anti-TB treatment was mainly because of nonviable MTB bacilli or NTM in patients with drug-susceptible pulmonary TB. AFB smear alone should not be used to assess treatment failure and careful examination of microbiologic status, including culture and drug susceptibility testing, is needed before making changes to retreatment regimens or empirical second

  13. Clinical significance of smear positivity for acid-fast bacilli after ≥5 months of treatment in patients with drug-susceptible pulmonary tuberculosis.

    PubMed

    Kang, Hyung Koo; Jeong, Byeong-Ho; Lee, Hyun; Park, Hye Yun; Jeon, Kyeongman; Huh, Hee Jae; Ki, Chang-Seok; Lee, Nam Yong; Koh, Won-Jung

    2016-08-01

    Patients with pulmonary tuberculosis (TB) with acid-fast bacilli (AFB)-positive sputum smear at 5 months or later during treatment are considered to be cases of treatment failure according to World Health Organization guidelines. This study evaluated the proportion, clinical characteristics, and significance of positive sputum smears after ≥5 months of standard treatment in patients with drug-susceptible pulmonary TB.This was a retrospective cohort study of 1611 patients with culture-confirmed drug-susceptible pulmonary TB who received standard anti-TB treatment from January 2009 to February 2014. Forty-one patients (2.5%) who were smear-positive after ≥5 months of treatment and 123 age- and sex-matched control patients were evaluated.Among the 41 smear-positive patients, culture of the sputum specimens yielded Mycobacterium tuberculosis (MTB) in 1 patient (2.4%), nontuberculous mycobacteria (NTM) in 7 (17.1%), and no growth in the remaining 33 patients (80.5%). Treatment was successfully completed in 40 patients (97.6%) with prolongation of the continuation phase regimens without change to second-line anti-TB treatment. In patients with smear positivity after ≥5 months of treatment compared with controls, cavitation on chest radiographs (53.7% vs. 25.2%, P = 0.001), bilateral involvement (51.2% vs. 30.1%, P = 0.01) and combined pleural effusion (26.8% vs. 10.6%, P = 0.01) were found more frequently at the time of treatment initiation, and paradoxical response occurred more commonly (19.5% vs. 3.3%, P = 0.002) during treatment.Smear-positive sputum after ≥5 months of standard anti-TB treatment was mainly because of nonviable MTB bacilli or NTM in patients with drug-susceptible pulmonary TB. AFB smear alone should not be used to assess treatment failure and careful examination of microbiologic status, including culture and drug susceptibility testing, is needed before making changes to retreatment regimens or empirical second-line anti

  14. Prevalence of smear-positive pulmonary tuberculosis among prisoners in Malawi: a national survey.

    PubMed

    Banda, H T; Gausi, F; Harries, A D; Salaniponi, F M

    2009-12-01

    A cross-sectional cell-to-cell survey was conducted in 18 of 22 prisons in Malawi to determine the period prevalence of smear-positive pulmonary tuberculosis (PTB). In each prison, prisoners were interviewed using a structured questionnaire. Prisoners with cough of >1 week's duration were investigated by sputum smear examination. Of 7661 prisoners, 3887 had cough of > or =1 week, of whom 3794 submitted three sputum specimens: 54 (0.7%) had smear-positive PTB. The prevalence of PTB was higher in large urban prisons (1.1%) than in district prisons (0.3%, P < 0.001). More needs to be done to improve TB control in urban prisons.

  15. COBAS® TaqMan® MTB, smear positivity grade and MGIT culture; correlation analyses of three methods for bacillary quantification.

    PubMed

    Chikamatsu, Kinuyo; Aono, Akio; Kato, Tomoko; Takaki, Akiko; Yamada, Hiroyuki; Sasaki, Yuka; Izumi, Kiyohiko; Yi, Lina; Mitarai, Satoshi

    2016-01-01

    We investigated the correlation between the cycle threshold (Ct) value of the COBAS(®) TaqMan(®) MTB (TaqMan MTB), the mycobacterial smear positivity grade, and the time to detection (TTD) in the Mycobacteria Growth Indicator Tube (MGIT) for quantification of Mycobacterium tuberculosis (MTB). For 57 sputum samples, significant correlations were observed between the Ct value, the smear positivity grade, and the MGIT TTD (Spearman's rank correlation coefficient: r(s) = -0.940, P < 0.001 and Pearson's correlation coefficient: r(p) = 0.737, P < 0.001). In addition, a correlation was observed between the number of bacteria estimated based on the smear positivity grade and the number of MTB bacilli calculated by the Ct value (r(s) = 0.930, P < 0.001). This study has demonstrated the possible estimation of the smear positivity grade and MGIT TTD using the Ct value of TaqMan MTB, which is based on a real-time PCR system, for diagnostic samples.

  16. LED fluorescence microscopy increases the detection of smear-positive pulmonary tuberculosis in medical colleges of India.

    PubMed

    Reza, L W; Satyanarayana, S; Pandey, A; Kumar, S; Devendrappa, N M; Anand, L; Singh, G; Kumar, A M V; Chadha, S S; Wilson, N; Sachdeva, K S; Nair, S A

    2013-09-21

    In July 2012, light-emitting diode fluorescence microscopy (LED-FM) replaced conventional light microscopy using Ziehl-Neelsen stain in the detection of sputum-positive pulmonary tuberculosis in 190 microscopy centres of medical colleges operating under India's Revised National Tuberculosis Control Programme. We compared the performance of LED-FM (July-December 2012) to that of conventional microscopy (July-December 2011) across 190 sites. Of 222 658 patients examined using conventional microscopy, 28 042 (12.6%) were smear-positive, while of 224 714 examined using LED-FM, 33 552 (14.9%) were smear-positive, an additional yield of 5251 cases after adjusting for the increase in patients examined. We recommend replacing conventional microscopy with LED-FM in high workload microscopy centres in India.

  17. Treatment outcome of new smear positive pulmonary tuberculosis patients in Penang, Malaysia

    PubMed Central

    2014-01-01

    included high-grade sputum and presence of lung cavities at the start of treatment, being alcoholic and elderly. Likewise, concurrent diabetes, presence of lung cavities at the start of the treatment and being a smoker were the significant predictors of longer treatment duration. Conclusion Our findings indicated that the treatment success rate among the new smear positive PTB patients was less than the success target set by World Health Organization. The proportion of patients in the successful outcome category may be increased by closely monitoring the treatment progress of the patients with aforementioned high risk characteristics. Similarly, more aggressive follow-up of the treatment defaulters and transferred out patients could also improve the TB treatment success rate. PMID:25037452

  18. Predicting the Incidence of Smear Positive Tuberculosis Cases in Iran Using Time Series Analysis

    PubMed Central

    MOOSAZADEH, Mahmood; KHANJANI, Narges; NASEHI, Mahshid; BAHRAMPOUR, Abbas

    2015-01-01

    Background: Determining the temporal variation and forecasting the incidence of smear positive tuberculosis (TB) can play an important role in promoting the TB control program. Its results may be used as a decision-supportive tool for planning and allocating resources. The present study forecasts the incidence of smear positive TB in Iran. Methods: This a longitudinal study using monthly tuberculosis incidence data recorded in the Iranian National Tuberculosis Control Program. The sum of registered cases in each month created 84 time points. Time series methods were used for analysis. Based on the residual chart of ACF, PACF, Ljung-Box tests and the lowest levels of AIC and BIC, the most suitable model was selected. Results: From April 2005 until March 2012, 34012 smear positive TB cases were recorded. The mean of TB monthly incidence was 404.9 (SD=54.7). The highest number of cases was registered in May and the difference in monthly incidence of smear positive TB was significant (P<0.001). SARIMA (0,1,1)(0,1,1)12 was selected as the most adequate model for prediction. It was predicted that the incidence of smear positive TB for 2015 will be about 9.8 per 100,000 people. Conclusion: Based on the seasonal pattern of smear positive TB recorded cases, seasonal ARIMA model was suitable for predicting its incidence. Meanwhile, prediction results show an increasing trend of smear positive TB cases in Iran. PMID:26744711

  19. Impact of changing diagnostic criteria for smear-positive tuberculosis: a cohort study in Malawi.

    PubMed

    Koole, O; Munthali, L; Mhango, B; Mpunga, J; Glynn, J R; Crampin, A C

    2014-07-01

    We assessed the impact on measured burden and outcomes of the revised World Health Organization and Malawi guidelines reclassifying people with single (including 'scanty') positive smears as smear-positive pulmonary tuberculosis cases. In a retrospective cohort in rural Malawi, 567 (34%) of 1670 smear-positive episodes were based on single positive smears (including 176 with scanty smears). Mortality rates and the proportion starting treatment were similar in those with two positive smears or single, non-scanty smears. Those with single scanty smears had higher mortality and a lower proportion starting treatment. The reclassification will increase the reported burden substantially, but should improve treatment access.

  20. Spatio-Temporal Analysis of Smear-Positive Tuberculosis in the Sidama Zone, Southern Ethiopia

    PubMed Central

    Dangisso, Mesay Hailu; Datiko, Daniel Gemechu; Lindtjørn, Bernt

    2015-01-01

    Background Tuberculosis (TB) is a disease of public health concern, with a varying distribution across settings depending on socio-economic status, HIV burden, availability and performance of the health system. Ethiopia is a country with a high burden of TB, with regional variations in TB case notification rates (CNRs). However, TB program reports are often compiled and reported at higher administrative units that do not show the burden at lower units, so there is limited information about the spatial distribution of the disease. We therefore aim to assess the spatial distribution and presence of the spatio-temporal clustering of the disease in different geographic settings over 10 years in the Sidama Zone in southern Ethiopia. Methods A retrospective space–time and spatial analysis were carried out at the kebele level (the lowest administrative unit within a district) to identify spatial and space-time clusters of smear-positive pulmonary TB (PTB). Scan statistics, Global Moran’s I, and Getis and Ordi (Gi*) statistics were all used to help analyze the spatial distribution and clusters of the disease across settings. Results A total of 22,545 smear-positive PTB cases notified over 10 years were used for spatial analysis. In a purely spatial analysis, we identified the most likely cluster of smear-positive PTB in 192 kebeles in eight districts (RR= 2, p<0.001), with 12,155 observed and 8,668 expected cases. The Gi* statistic also identified the clusters in the same areas, and the spatial clusters showed stability in most areas in each year during the study period. The space-time analysis also detected the most likely cluster in 193 kebeles in the same eight districts (RR= 1.92, p<0.001), with 7,584 observed and 4,738 expected cases in 2003-2012. Conclusion The study found variations in CNRs and significant spatio-temporal clusters of smear-positive PTB in the Sidama Zone. The findings can be used to guide TB control programs to devise effective TB control

  1. True status of smear-positive pulmonary tuberculosis defaulters in Malawi.

    PubMed Central

    Kruyt, M. L.; Kruyt, N. D.; Boeree, M. J.; Harries, A. D.; Salaniponi, F. M.; van Noord, P. A.

    1999-01-01

    The article reports the results of a study to determine the true outcome of 8 months of treatment received by smear-positive pulmonary tuberculosis (PTB) patients who had been registered as defaulters in the Queen Elizabeth Central Hospital (QECH) and Mlambe Mission Hospital (MMH), Blantyre, Malawi. The treatment outcomes were documented from the tuberculosis registers of all patients registered between 1 October 1994 and 30 September 1995. The true treatment outcome for patients who had been registered as defaulters was determined by making personal inquiries at the treatment units and the residences of patients or relatives and, in a few cases, by writing to the appropriate postal address. Interviews were carried out with patients who had defaulted and were still alive and with matched, fully compliant PTB patients who had successfully completed the treatment to determine the factors associated with defaulter status. Of the 1099 patients, 126 (11.5%) had been registered as defaulters, and the true treatment outcome was determined for 101 (80%) of the latter; only 22 were true defaulters, 31 had completed the treatment, 31 had died during the treatment period, and 17 had left the area. A total of 8 of the 22 true defaulters were still alive and were compared with the compliant patients. Two significant characteristics were associated with the defaulters; they were unmarried; and they did not know the correct duration of antituberculosis treatment. Many of the smear-positive tuberculosis patients who had been registered as defaulters in the Blantyre district were found to have different treatment outcomes, without defaulting. The quality of reporting in the health facilities must therefore be improved in order to exclude individuals who are not true defaulters. PMID:10361755

  2. True status of smear-positive pulmonary tuberculosis defaulters in Malawi.

    PubMed

    Kruyt, M L; Kruyt, N D; Boeree, M J; Harries, A D; Salaniponi, F M; van Noord, P A

    1999-01-01

    The article reports the results of a study to determine the true outcome of 8 months of treatment received by smear-positive pulmonary tuberculosis (PTB) patients who had been registered as defaulters in the Queen Elizabeth Central Hospital (QECH) and Mlambe Mission Hospital (MMH), Blantyre, Malawi. The treatment outcomes were documented from the tuberculosis registers of all patients registered between 1 October 1994 and 30 September 1995. The true treatment outcome for patients who had been registered as defaulters was determined by making personal inquiries at the treatment units and the residences of patients or relatives and, in a few cases, by writing to the appropriate postal address. Interviews were carried out with patients who had defaulted and were still alive and with matched, fully compliant PTB patients who had successfully completed the treatment to determine the factors associated with defaulter status. Of the 1099 patients, 126 (11.5%) had been registered as defaulters, and the true treatment outcome was determined for 101 (80%) of the latter; only 22 were true defaulters, 31 had completed the treatment, 31 had died during the treatment period, and 17 had left the area. A total of 8 of the 22 true defaulters were still alive and were compared with the compliant patients. Two significant characteristics were associated with the defaulters; they were unmarried; and they did not know the correct duration of antituberculosis treatment. Many of the smear-positive tuberculosis patients who had been registered as defaulters in the Blantyre district were found to have different treatment outcomes, without defaulting. The quality of reporting in the health facilities must therefore be improved in order to exclude individuals who are not true defaulters.

  3. Adverse Events in Treating Smear-Positive Tuberculosis Patients in China.

    PubMed

    Zhang, Tao; Du, Jian; Yin, Xiaoyan; Xue, Fuzhong; Liu, Yanxun; Li, Runzi; Luo, Cheng; Li, Liang; Li, Xiujun

    2016-01-01

    This study aimed to estimate the adverse events (AE) rate during anti-tuberculosis treatment and to explore AE-related risk factors. New and previously treated smear-positive tuberculosis (TB) cases were enrolled from eight regions in China between April 2009 and October 2010. The AE rate was estimated, and AE risk factors during anti-TB treatment were assessed using Cox proportional models. Among 2091 Chinese subjects with anti-TB treatment, 462 (22.1%, 95% confidence interval (CI), 20.3-23.9) patients developed AE, with liver injury and gastrointestinal reactions constituting the most common AE. Specifically, 9.8% (95% CI, 8.5-11.1) and 6.3% (95% CI, 5.3-7.4) developed liver injuries and gastrointestinal reactions, respectively. We found that AE rate differed by regions, TB knowledge score, symptoms score and smoking status. Liver injuries were associated with age, sex and smoking status; gastrointestinal reactions were associated with education level and symptom score. Improving patients' knowledge on TB could reduce AE rate.

  4. Comparison of two active case-finding strategies for community-based diagnosis of symptomatic smear-positive tuberculosis and control of infectious tuberculosis in Harare, Zimbabwe (DETECTB): a cluster-randomised trial

    PubMed Central

    Corbett, Elizabeth L; Bandason, Tsitsi; Duong, Trinh; Dauya, Ethel; Makamure, Beauty; Churchyard, Gavin J; Williams, Brian G; Munyati, Shungu S; Butterworth, Anthony E; Mason, Peter R; Mungofa, Stanley; Hayes, Richard J

    2010-01-01

    Summary Background Control of tuberculosis in settings with high HIV prevalence is a pressing public health priority. We tested two active case-finding strategies to target long periods of infectiousness before diagnosis, which is typical of HIV-negative tuberculosis and is a key driver of transmission. Methods Clusters of neighbourhoods in the high-density residential suburbs of Harare, Zimbabwe, were randomised to receive six rounds of active case finding at 6-monthly intervals by either mobile van or door-to-door visits. Randomisation was done by selection of discs of two colours from an opaque bag, with one disc to represent every cluster, and one colour allocated to each intervention group before selection began. In both groups, adult (≥16 years) residents volunteering chronic cough (≥2 weeks) had two sputum specimens collected for fluorescence microscopy. Community health workers and cluster residents were not masked to intervention allocation, but investigators and laboratory staff were masked to allocation until final analysis. The primary outcome was the cumulative yield of smear-positive tuberculosis per 1000 adult residents, compared between intervention groups; analysis was by intention to treat. The secondary outcome was change in prevalence of culture-positive tuberculosis from before intervention to before round six of intervention in 12% of randomly selected households from the two intervention groups combined; analysis was based on participants who provided sputum in the two prevalence surveys. This trial is registered, number ISRCTN84352452. Findings 46 study clusters were identified and randomly allocated equally between intervention groups, with 55 741 adults in the mobile van group and 54 691 in the door-to-door group at baseline. HIV prevalence was 21% (1916/9060) and in the 6 months before intervention the smear-positive case notification rate was 2·8 per 1000 adults per year. The trial was completed as planned with no adverse events

  5. A Comparative Study of Induced Sputum and Bronchial Washings in Diagnosing Sputum Smear Negative Pulmonary Tuberculosis

    PubMed Central

    Mandava, Venu; Namballa, Usha Rani; Makala, Sravani

    2016-01-01

    Introduction Tuberculosis is one of the most important public health problem worldwide. Detecting patients with active pulmonary Koch’s disease is an important component of tuberculosis control programs. However, at times in patients even with a compatible clinical picture, sputum smears do not reveal acid-fast bacilli and smear-negative pulmonary tuberculosis remains a common problem. Aim The present study is aimed to compare the results of induced sputum and bronchial washings smear in patients suspected to have sputum smear negative pulmonary tuberculosis. Materials and Methods A prospective study conducted from August 2014 to July 2015, comprising 120 patients fulfilling study criteria. Patients with respiratory symptoms and chest roentgenogram suspicious of pulmonary tuberculosis with no previous history of anti-tuberculosis treatment and two spontaneous sputum smear samples negative for acid fast bacilli were included. Patients with active haemoptysis and sputum positive were excluded from the study. Sputum induction was done by using 5-10 ml of 3% hypertonic saline through ultrasonic nebulizer taking safety precautions. All the patient underwent fibreoptic bronchoscopy after six hours fasting on the same day. About 20 ml of normal saline instilled into the suspected pathology area and washings were taken with gentle suction. The sample processing and fluorescent staining for acid fast bacilli was done in a designated microscopy lab. Results Out of 120 sputum smear negative pulmonary tuberculosis patients, induced sputum smear examination detected acid fast bacilli in 76 patients (63.3%) and acid fast bacilli detected from bronchial washings in 94 patients (78.3%). Smear positivity was higher in cavitary and infiltrative lesions as compared to consolidation and infrahilar pattern disease. Conclusion Even though both induced sputum and bronchial washings procedures were valuable for the diagnosis of sputum smear negative, sputum induction with hypertonic

  6. Impact of Educational Intervention on Patients Behavior with Smear-positive Pulmonary Tuberculosis: A Study Using the Health Belief Model

    PubMed Central

    Jadgal, Khair Mohammad; Nakhaei-Moghadam, Tayebeh; Alizadeh-Seiouki, Hadi; Zareban, Iraj; Sharifi-Rad, Javad

    2015-01-01

    Introduction: Tuberculosis is a single-agent infectious disease, which is the major cause of death around the world. Approximately one third of the world’s population is infected with tuberculosis (TB) bacilli and at risk of developing active TB. The purpose of this study was determined the impact of education based on health belief model in promoting behavior of smear-positive pulmonary TB among patients in Chabahar city, Iran. Material and methods: Of the 80 smear-positive pulmonary TB who referred to health centers in Chabahar voluntarily participated in this interventional study. The data collected using questionnaire based on health belief model. The data were analyzed by using paired t-test, independent t-test, pearson correlation and chi-square test with SPSS 16. Results: The cognitive skills were increased significantly from 6.10 to 6.88 after intervention. All behavioral skills were increased significantly from 2.08 to 2.88 after implementing the intervention. Perceived severity was increased from11.08to12.19 significantly. Percepted benefits were enhanced significantly from 11.48 to 12.23. Mean percepted barrier was decreased significantly from 17.52 to 16.68. Conclusion: Findings demonstrated that implementing educational intervention programs can increase the level of knowledge and behavior of patients regarding smear- positive pulmonary TB initiatives. PMID:26543411

  7. Frequency of Latent and Smear Positive Tuberculosis in Chronic Psychotic Disorders

    PubMed Central

    Ebrahimi, Hannan; Mohammadi, Alieh; Jahromi, Sina Khajeh; Naghdipour, Misa; Ebrahimi, Hossein

    2013-01-01

    Objective Screening is one of the ways to combat Tuberculosis (TB) and should be mostly concentrated on groups showing some symptoms of the disease. Tuberculosis can be transferred from person to person in laboratories, prisons and psychiatry hospitals. The purpose of this study was to survey pulmonary TB in patients with schizophrenia in Rasht. Methods In this descriptive-cross sectional, Two hundred fifty seven consecutive patients with chronic psychotic disorder hospitalized in psychotic hospitals underwent purified protein derivative (PPD) test. PPD test was done with the unit 5T which was injected subcutaneously on anterior surface and at the top of left forearm. The results of the test were interpreted by the pen technique method and based on transverse diameter of induration of about 48-72 hrs. Induration size due to hypersensitivity to PPD more than 10mm was considered positive. Patients with positive PPD test underwent complementary sputum smear. Data were analyzed using chi- square and T test. Results The mean age of patients was 45±10 years; 75.5% were male, 74.7% were single, 10.5% married, 7.8% divorced, and 68.1% were smokers. These patients suffered from chronic psychotic disorder at the mean time of 15±7.9 years. In 74 patients (28%) positive PPD test were recorded, but active pulmonary TB was not found in complementary experiments of PPD sample. Based on data analysis, only age and gender showed a significant relationship with the results of the PPD test (P < 0.05). Conclusions This study showed that patients with positive PPD test are much more than the normal population, but active pulmonary TB was not observed in our samples. Since these patients are in clinical and closed places, more programs for screening are required. PMID:23682252

  8. Determination of mycobacterial antigens in sputum by enzyme immunoassay.

    PubMed Central

    Yáñez, M A; Coppola, M P; Russo, D A; Delaha, E; Chaparas, S D; Yeager, H

    1986-01-01

    An enzyme-linked immunosorbent assay (ELISA) was examined for its usefulness in detecting mycobacterial antigens in sputum. A double-antibody sandwich procedure was set up by using a commercially available hyperimmune serum directed against Mycobacterium bovis, BCG. The ELISA was able to detect 10 ng of protein per ml of BCG sonic extract. The system also clearly distinguished Mycobacterium tuberculosis organisms from Mycobacterium avium and Mycobacterium kansasii organisms. A total of 68 unknown sputum specimens submitted to the clinical laboratories for examination for tuberculosis were tested by ELISA. Of the 20 specimens that were smear positive and culture positive, 12 (60%) were positive by ELISA; 6 of the 11 (55%) smear-positive culture-negative samples were positive by ELISA; 1 of 2 (50%) of the smear-negative culture-positive samples was positive by ELISA; and only 3 of 35 (9%) of the smear-negative culture-negative samples were positive by ELISA. This approach offers promise as an aid in the presumptive differentiation of nontuberculous mycobacteria from the M. tuberculosis complex. PMID:3086369

  9. Did FIDELIS projects contribute to the detection of new smear-positive pulmonary tuberculosis cases in China?

    PubMed Central

    Rusen, I. D.; Hinderaker, S. G.; Roldan, A.; Heldal, E.; Enarson, D. A.; Zhang, L-X.

    2016-01-01

    Setting: The first phase of the Fund for Innovative DOTS Expansion through Local Initiatives to Stop TB (FIDELIS) projects in China started in 2003. Objective: To determine whether the FIDELIS projects contributed to the increased case detection rate for new smear-positive pulmonary tuberculosis (PTB) in China. Methods: We compared the case notification rates (CNRs) in the intervention year with those of the previous year in the FIDELIS areas, then compared the difference between the CNRs of the intervention year and the previous year in the FIDELIS areas with those in the non-FI-DELIS areas within the province. Results: There was an increase in the CNR in the intervention year compared with the previous year for all the project sites. The differences between the CNR in the intervention year and the previous year ranged from 6.4 to 31.1 per 100 000 population in the FIDELIS areas and from 2.9 to 20.4/100 000 in the non-FIDELIS areas. Differences-in-differences analysis shows that the differences in the CNRs in the FIDELIS areas were not statistically significantly different from those in the non-FIDELIS areas (P = 0.393). Conclusion: The FIDELIS projects may have contributed to the increase in case detection of new smear-positive PTB in China, but the level of evidence is low. PMID:27695680

  10. Did FIDELIS projects contribute to the detection of new smear-positive pulmonary tuberculosis cases in China?

    PubMed Central

    Rusen, I. D.; Hinderaker, S. G.; Roldan, A.; Heldal, E.; Enarson, D. A.; Zhang, L-X.

    2016-01-01

    Setting: The first phase of the Fund for Innovative DOTS Expansion through Local Initiatives to Stop TB (FIDELIS) projects in China started in 2003. Objective: To determine whether the FIDELIS projects contributed to the increased case detection rate for new smear-positive pulmonary tuberculosis (PTB) in China. Methods: We compared the case notification rates (CNRs) in the intervention year with those of the previous year in the FIDELIS areas, then compared the difference between the CNRs of the intervention year and the previous year in the FIDELIS areas with those in the non-FI-DELIS areas within the province. Results: There was an increase in the CNR in the intervention year compared with the previous year for all the project sites. The differences between the CNR in the intervention year and the previous year ranged from 6.4 to 31.1 per 100 000 population in the FIDELIS areas and from 2.9 to 20.4/100 000 in the non-FIDELIS areas. Differences-in-differences analysis shows that the differences in the CNRs in the FIDELIS areas were not statistically significantly different from those in the non-FIDELIS areas (P = 0.393). Conclusion: The FIDELIS projects may have contributed to the increase in case detection of new smear-positive PTB in China, but the level of evidence is low.

  11. Trend of Smear-positive Pulmonary Tuberculosis in Iran during 1995–2012: A Segmented Regression Model

    PubMed Central

    Khazaei, Salman; Soheilyzad, Mokhtar; Molaeipoor, Leila; Khazaei, Zaher; Rezaeian, Shahab; Khazaei, Somayeh

    2016-01-01

    Background: Describing trend in tuberculosis (TB) over time can play an important role to assess the disease control strategies and predict the future morbidity and mortality. This study aimed to determine the incidence trend of smear-positive pulmonary tuberculosis (SPPT) in sub-age and sex groups during the years of 1995–2012. Methods: This retrospective cohort study was performed in 2015 by using the dataset regarding National Statistics of SPPT reported by World Health Organization during 1995–2012. Annual percent changes (APCs) and average annual percent changes (AAPCs) were estimated to determine the summery statistics of trend using segmented regression model. Results: During 1995–2012, there were 96,579 SPPT case notifications in Iran (male to female ratio: 0.99). There was only one change point in 1997 for SPPT incidence in subgroups of age and sex during 1995–2012. The AAPCs for both genders and also all three age groups had a significant descending trend during the time period (P < 0.05). Conclusions: Our results showed a downward trend in the SPPT incidence. It seems that to achieve the set goals and high successful in TB control program especially reduction in SPPT, pay more attention to old age and males should be considered. In addition, improvement of clinical and medical care services and notification processes would be imperative. PMID:27413517

  12. Routine sputum culture

    MedlinePlus

    Sputum culture ... There, it is placed in a special dish (culture). It is then watched to see if bacteria ... Chernecky CC, Berger BJ. Culture, routine. In: Chernecky CC, Berger BJ, ... . 6th ed. Philadelphia, PA: Elsevier Saunders; 2013:409- ...

  13. DIAGNOSTIC TEST OF SPUTUM GENEXPERT MTB/RIF FOR SMEAR NEGATIVE PULMONARY TUBERCULOSIS.

    PubMed

    Reechaipichitkul, Wipa; Phetsuriyawong, Attawoot; Chaimanee, Prajuab; Ananta, Pimjai

    2016-05-01

    The objective of this study was to evaluate the performance of the Gene-Xpert MTB/RIF sputum test for diagnosing pulmonary tuberculosis (TB) among patients sputum acid-fast bacillus (AFB) smear negative results in Thailand, a country with a high prevalence of pulmonary tuberculosis. We studied 151 patients who presented to Srinagarind Hospital, Khon Kaen, Thailand with a 2 week or more history of fever and/or cough and an abnormal chest radiograph between 2010 and 2014; these patients had at least 2 negative sputum AFB smear results. Of these, 76 were diagnosed as having either confirmed or probable pulmonary TB: the 32 confirmed cases were those with a positive sputum culture for Mycobacterium tuberculosis (MTB) and the 44 probable case were those with clinical and radiographic findings consistent with TB and who had a response to anti-TB therapy. Seventy-five cases were diagnosed as not having pulmonary TB. Of the 32 patients with a positive sputum culture for MTB, 26 had a positive GeneXpert MTB/RIF sputum test. Compared to sputum culture for MTB the GeneXpert MTB/ RIF test gave a sensitivity of 83.9% (95% CI: 66.3-94.5) and a specificity of 92.1% (95% CI: 83.6-97), a positive predictive value (PPV) of 81.3% (95% CI: 63.6-92.8) and a negative predictive value (NPV) of 93.3% (95% CI: 85.1-97.8). The GeneXpert MTB/RIF test had a fair sensitivity and specificity for diagnosing smear negative pulmonary TB. It may be useful for diagnosing pulmonary TB in patients with a negative sputum AFB smear. The assay is faster than culture and can detect rifampicin resistant strains of MTB. PMID:27405129

  14. Black-pigmented sputum

    PubMed Central

    Martínez-Girón, Rafael; Mosquera-Martínez, Joaquín; Martínez-Torre, Santiago

    2013-01-01

    Black-pigmented sputum, also called “melanoptysis,” is a symptom that may be observed in certain pathologies such us coal workers’ pneumoconiosis (anthracosis). The cavitation and liquefaction of a fibrosed mass by an infectious process (tuberculosis, infections by anaerobes, etc.) or by ischemic necrosis may cause expectoration of a blackish secretion. We report the case of a patient with labor precedents as a coal worker, from whom abundant black sputum was obtained in the course of an incidental expectoration. Cyto-histological findings are shown and a differential diagnosis is established. PMID:24648674

  15. The effectiveness of sputum pH analysis in the prediction of response to therapy in patients with pulmonary tuberculosis.

    PubMed

    Masuda, Makoto; Sato, Takashi; Sakamaki, Kentaro; Kudo, Makoto; Kaneko, Takeshi; Ishigatsubo, Yoshiaki

    2015-01-01

    Purpose. The predictive factor of response to antituberculous therapy has not been fully elucidated. Airway acidity has been thought to be a potential indicator of the bactericidal activity. Therefore, we hypothesized that monitoring airway acidity by measuring sputum pH could predict response to therapy. Methods. A total of 47 patients having newly diagnosed, smear-positive, active pulmonary tuberculosis were enrolled between October 2011 and March 2014. Sputum samples were serially analyzed before and after treatment. Eligible patients who initiated a standard 6-month treatment were monitored for the length of time to sputum smear and culture conversion. Results. There were 39 patients who completed a 2-month intensive phase of isoniazid, rifampicin, pyrazinamide, and ethambutol therapy followed by a 4-month continuation phase of isoniazid and rifampicin. Although factors including age, cavitation, sputum grade, and use of an acid-suppressant were associated with initial low sputum pH in univariate analysis, multivariate analysis revealed that only age ≥61 years was a statistically important factor predicting low pH value (p = 0.005). Further outcome analysis showed that initial low sputum pH before treatment was the only factor significantly associated with shorter length of time to both sputum smear and culture conversion (p = 0.034 and 0.019, respectively) independent of the effects of age, sputum bacterial load, extent of lung lesion, and cavitation. Thus, initial low sputum pH indicated favorable response to anti-tuberculosis therapy. Conclusions. Measuring sputum pH is an easy and inexpensive way of predicting response to standard combination therapy in patients with pulmonary tuberculosis. PMID:26644982

  16. Griffiss AFB integrated resource assessment

    SciTech Connect

    Dixon, D.R.; Armstrong, P.R.; Keller, J.M.

    1993-02-01

    The US Air Force Air Combat Command has tasked the Pacific Northwest Laboratory (PNL) as the lead laboratory supporting the US Department of Energy (DOE) Federal Energy Management Program's (FEMP) mission to identify, evaluate, and assist in acquiring all cost-effective energy projects at Griffiss Air Force Base (AFB). This is a model program PNL is designing for federal customers served by the Niagara Mohawk Power Company (Niagara Mohawk). It will (1) identify and evaluate all electric cost-effective energy projects; (2) develop a schedule at each installation for project acquisition considering project type, size, timing, and capital requirements, as well as energy and dollar savings; and (3) secure 100% of the financing required to implement electric energy efficiency projects from Niagara Mohawk and have Niagara Mohawk procure the necessary contractors to perform detailed audits and install the technologies. This report documents the assessment of baseline energy use at one of Niagara Mohawk's primary federal facilities, Griffiss AFB, an Air Combat Command facility located near Rome, New York. It is a companion report to Volume 1, the Executive Summary, and Volume 3, the Electric Resource Assessment. The analysis examines the characteristics of electric, gas, oil, propane, coal, and purchased thermal capacity use for fiscal year (FY) 1990. The results include energy-use intensities for the facilities at Griffiss AFB by building type and electric energy end use. A complete electric energy consumption reconciliation is presented that accounts for the distribution of all major electric energy uses and losses among buildings, utilities, and central systems.

  17. Anisotropic tubular filtering for automatic detection of acid-fast bacilli in Ziehl-Neelsen stained sputum smear samples

    NASA Astrophysics Data System (ADS)

    Raza, Shan-e.-Ahmed; Marjan, M. Q.; Arif, Muhammad; Butt, Farhana; Sultan, Faisal; Rajpoot, Nasir M.

    2015-03-01

    One of the main factors for high workload in pulmonary pathology in developing countries is the relatively large proportion of tuberculosis (TB) cases which can be detected with high throughput using automated approaches. TB is caused by Mycobacterium tuberculosis, which appears as thin, rod-shaped acid-fast bacillus (AFB) in Ziehl-Neelsen (ZN) stained sputum smear samples. In this paper, we present an algorithm for automatic detection of AFB in digitized images of ZN stained sputum smear samples under a light microscope. A key component of the proposed algorithm is the enhancement of raw input image using a novel anisotropic tubular filter (ATF) which suppresses the background noise while simultaneously enhancing strong anisotropic features of AFBs present in the image. The resulting image is then segmented using color features and candidate AFBs are identified. Finally, a support vector machine classifier using morphological features from candidate AFBs decides whether a given image is AFB positive or not. We demonstrate the effectiveness of the proposed ATF method with two different feature sets by showing that the proposed image analysis pipeline results in higher accuracy and F1-score than the same pipeline with standard median filtering for image enhancement.

  18. ASSESSMENT OF DETECTION EFFICACY OF MYCOBACTERIUM TUBERCULOSIS IN SPUTUM SAMPLES BY REAL TIME PCR BASED METHOD.

    PubMed

    Upadhyaya, S K; Dixit, S M; Shrestha, S; Dangol, S D; Pokhrel, D; Banjara, S; Shrestha, K S; Hengoju, S; Dahal, B K

    2013-07-01

    Tuberculosis (TB) is a major public health problem in Nepal and ranks as one of the most prevalent communicable diseases throughout the country. In Nepal, 45% of total population is infected with TB and 40,000 people get TB every year. Twenty thousand new sputum positive cases are seen every year and 5000-7000 people die each year from TB. Thirty sputum samples were collected from Sukraraj Tropical and Infectious Disease Hospital, Teku, Kathmandu, Nepal and the comparative study of Acid-fast Bacilli (AFB) test and Real time PCR were conducted separately with the culture test which is regarded as gold standard by WHO. Detection of Mycobacterium tuberculosis through use of Real time PCR was found to be higher as compared to AFB and culture. Real-time PCR test showed higher sensitivity (100%) and specificity (94.11%) as compared to AFB test with sensitivity of 84.61% and specificity of 88.24%. Positive predictive value was found to be 84.61% and 92.86% for AFB and Q-PCR respectively. Negative predictive value was found to be 88.24%, and for Q-PCR, it was found to be 100%. Our statistics clearly show that TB diagnosis by Q-PCR is highly efficient and reliableover conventional methods of diagnosis and here we recommend its use in the hospitals and clinics of Nepal.

  19. Adherence to Tuberculosis Treatment, Sputum Smear Conversion and Mortality: A Retrospective Cohort Study in 48 Rwandan Clinics

    PubMed Central

    Kayigamba, Felix R.; Bakker, Mirjam I.; Mugisha, Veronicah; De Naeyer, Ludwig; Gasana, Michel; Cobelens, Frank; van der Loeff, Maarten Schim

    2013-01-01

    Background Adherence to treatment and sputum smear conversion after 2 months of treatment are thought to be important for successful outcome of tuberculosis (TB) treatment. Methods Retrospective cohort study of new adult TB patients diagnosed in the first quarter of 2007 at 48 clinics in Rwanda. Data were abstracted from TB registers and individual treatment charts. Logistic regression analysis was done to examine associations between baseline demographic and clinical factors and three outcomes adherence, sputum smear conversion at two months, and death. Results Out of 725 eligible patients the treatment chart was retrieved for 581 (80%). Fifty-six (10%) of these patients took <90% of doses (defined as poor adherence). Baseline demographic characteristics were not associated with adherence to TB treatment, but adherence was lower among HIV patients not taking antiretroviral therapy (ART); p = 0.03). Sputum smear results around 2 months after start of treatment were available for 220 of 311 initially sputum-smear-positive pulmonary TB (PTB+) patients (71%); 175 (80%) had achieved sputum smear conversion. In multivariable analysis, baseline sputum smear grade (odds ratio [OR] = 2.7, 95% Confidence interval [CI] 1.1–6.6 comparing smear 3+ against 1+) and HIV infection (OR 3.0, 95%CI 1.3–6.7) were independent predictors for non-conversion at 2 months. Sixty-nine of 574 patients (12%) with known TB treatment outcomes had died. Besides other known determinants, poor adherence had an independent, strong effect on mortality (OR 3.4, 95%CI 1.4–7.8). Conclusion HIV infection is an important independent predictor of failure of sputum smear conversion at 2 months among PTB+ patients. Poor adherence to TB treatment is an important independent determinant of mortality. PMID:24066053

  20. The diagnostic accuracy of the MTBDRplus and MTBDRsl assays for drug-resistant TB detection when performed on sputum and culture isolates.

    PubMed

    Tomasicchio, Michele; Theron, Grant; Pietersen, Elize; Streicher, Elizabeth; Stanley-Josephs, Danielle; van Helden, Paul; Warren, Rob; Dheda, Keertan

    2016-01-01

    Although molecular tests for drug-resistant TB perform well on culture isolates, their accuracy using clinical samples, particularly from TB and HIV-endemic settings, requires clarification. The MTBDRplus and MTBDRsl line probe assays were evaluated in 181 sputum samples and 270 isolates from patients with culture-confirmed drug-sensitive-TB, MDR-TB, or XDR-TB. Phenotypic culture-based testing was the reference standard. Using sputum, the sensitivities for resistance was 97.7%, 95.4%, 58.9%, 61.6% for rifampicin, isoniazid, ofloxacin, and amikacin, respectively, whereas the specificities were 91.8%, 89%, 100%, and 100%, respectively. MTBDRsl sensitivity differed in smear-positive vs. smear-negative samples (79.2% vs. 20%, p < 0.0001 for ofloxacin; 72.9% vs. 37%, p = 0.0023 for amikacin) but not by HIV status. If used sequentially, MTBDRplus and MTBDRsl could rule-in XDR-TB in 78.5% (22/28) and 10.5% (2/19) of smear-positive and smear-negative samples, respectively. On culture isolates, the sensitivity for resistance to rifampicin, isoniazid, ofloxacin, and amikacin was 95.1%, 96.1%, 72.3% and 76.6%, respectively, whereas the specificities exceeded 96%. Using a sequential testing approach, rapid sputum-based diagnosis of fluoroquinolone or aminoglycoside-resistant TB is feasible only in smear-positive samples, where rule-in value is good. Further investigation is required in samples that test susceptible in order to rule-out second-line drug resistance. PMID:26860462

  1. Do Instructional Videos on Sputum Submission Result in Increased Tuberculosis Case Detection? A Randomized Controlled Trial

    PubMed Central

    Doulla, Basra; Mhimbira, Francis; Mtutu, Hawa; Hiza, Helen; Sasamalo, Mohamed; Rutaihwa, Liliana; Rieder, Hans L.; Seimon, Tamsyn; Mutayoba, Beatrice; Weiss, Mitchell G.; Fenner, Lukas

    2015-01-01

    Objective We examined the effect of an instructional video about the production of diagnostic sputum on case detection of tuberculosis (TB), and evaluated the acceptance of the video. Trial Design Randomized controlled trial. Methods We prepared a culturally adapted instructional video for sputum submission. We analyzed 200 presumptive TB cases coughing for more than two weeks who attended the outpatient department of the governmental Municipal Hospital in Mwananyamala (Dar es Salaam, Tanzania). They were randomly assigned to either receive instructions on sputum submission using the video before submission (intervention group, n = 100) or standard of care (control group, n = 100). Sputum samples were examined for volume, quality and presence of acid-fast bacilli by experienced laboratory technicians blinded to study groups. Results Median age was 39.1 years (interquartile range 37.0–50.0); 94 (47%) were females, 106 (53%) were males, and 49 (24.5%) were HIV-infected. We found that the instructional video intervention was associated with detection of a higher proportion of microscopically confirmed cases (56%, 95% confidence interval [95% CI] 45.7–65.9%, sputum smear positive patients in the intervention group versus 23%, 95% CI 15.2–32.5%, in the control group, p <0.0001), an increase in volume of specimen defined as a volume ≥3ml (78%, 95% CI 68.6–85.7%, versus 45%, 95% CI 35.0–55.3%, p <0.0001), and specimens less likely to be salivary (14%, 95% CI 7.9–22.4%, versus 39%, 95% CI 29.4–49.3%, p = 0.0001). Older age, but not the HIV status or sex, modified the effectiveness of the intervention by improving it positively. When asked how well the video instructions were understood, the majority of patients in the intervention group reported to have understood the video instructions well (97%). Most of the patients thought the video would be useful in the cultural setting of Tanzania (92%). Conclusions Sputum submission instructional videos increased

  2. Quantitation of Microorganisms in Sputum

    PubMed Central

    Monroe, P. W.; Muchmore, H. G.; Felton, F. G.; Pirtle, J. K.

    1969-01-01

    A method of quantitating microbial cultures of homogenized sputum has been devised. Possible application of this method to the problem of determining the etiologic agent of lower-respiratory-tract infections has been studied to determine its usefulness as a guide in the management of these infections. Specimens were liquefied by using an equal volume of 2% N-acetyl-L-cysteine. The liquefied sputum suspension was serially diluted to 10-1, 10-3, 10-5, and 10-7. These dilutions were plated on appropriate media by using an 0.01-ml calibrated loop; they were incubated and examined by standard diagnostic methods. Quantitation of fresh sputum from patients with pneumonia prior to antimicrobial therapy revealed that probable pathogens were present in populations of 107 organisms/ml or greater. Normal oropharyngeal flora did not occur in these numbers before therapy. Comparison of microbial counts on fresh and aged sputum showed that it is necessary to use only fresh specimens, since multiplication or death alters both quantitative and qualitative findings. Proper collection and quantitative culturing of homogenized sputum provided information more directly applicable to patient management than did qualitative routine methods. Not only was the recognition of the probable pathogenic organism in pneumonia patients improved, but serial quantitative cultures were particularly useful in recognizing the emergence of superinfections and in evaluating the efficacy of antimicrobial therapy. PMID:4390055

  3. The quality of sputum smear microscopy in public-private mix directly observed treatment laboratories in West Amhara region, Ethiopia.

    PubMed

    Manalebh, Almaw; Demissie, Meaza; Mekonnen, Daniel; Abera, Bayeh

    2015-01-01

    Ethiopia adopted Public-Private Mix Directly Observed Treatment Short Course Chemotherapy (PPM-DOTS) strategy for tuberculosis (TB) control program. Quality of sputum smear microscopy has paramount importance for tuberculosis control program in resource-poor countries like Ethiopia. A cross-sectional study was conducted to assess the quality of sputum smear microscopy in 37 Public-Private Mix laboratories in West Amhara, Ethiopia. The three external quality assessment methods (onsite evaluation, panel testing and blind rechecking) were employed. Onsite assessment revealed that 67.6% of PPM-DOTS laboratories were below the standard physical space (5 X 6) m2. The average monthly workload per laboratory technician was 19.5 (SD±2.9) slides with 12.8% positivity rate. The quality of Acid Fast Bacilli (AFB) staining reagents was sub-standard. The overall agreement for blind rechecking of 1,123 AFB slides was 99.4% (Kappa = 0.97). Reading of 370 AFB panel slides showed 3.5% false reading (Kappa = 0.92). Moreover, the consistency of reading scanty bacilli slides was lower (93%) compared to 1+, 2+ and 3+ bacilli. Based on blind rechecking and panel testing results, PPM-DOTS site laboratories showed good agreement with the reference laboratory. Physical space and qualities of AFB reagents would be areas of intervention to sustain the quality of sputum smear microscopy. Therefore, regular external quality assessment and provision of basic laboratory supplies for TB diagnosis would be the way forward to improve the quality of sputum smear microscopy services in PPM-DOTS laboratories.

  4. The Quality of Sputum Smear Microscopy in Public-Private Mix Directly Observed Treatment Laboratories in West Amhara Region, Ethiopia

    PubMed Central

    Manalebh, Almaw; Demissie, Meaza; Mekonnen, Daniel; Abera, Bayeh

    2015-01-01

    Ethiopia adopted Public-Private Mix Directly Observed Treatment Short Course Chemotherapy (PPM-DOTS) strategy for tuberculosis (TB) control program. Quality of sputum smear microscopy has paramount importance for tuberculosis control program in resource-poor countries like Ethiopia. A cross-sectional study was conducted to assess the quality of sputum smear microscopy in 37 Public-Private Mix laboratories in West Amhara, Ethiopia. The three external quality assessment methods (onsite evaluation, panel testing and blind rechecking) were employed. Onsite assessment revealed that 67.6% of PPM-DOTS laboratories were below the standard physical space (5 X 6) m2. The average monthly workload per laboratory technician was 19.5 (SD±2.9) slides with 12.8% positivity rate. The quality of Acid Fast Bacilli (AFB) staining reagents was sub-standard. The overall agreement for blind rechecking of 1,123 AFB slides was 99.4% (Kappa = 0.97). Reading of 370 AFB panel slides showed 3.5% false reading (Kappa = 0.92). Moreover, the consistency of reading scanty bacilli slides was lower (93%) compared to 1+, 2+ and 3+ bacilli. Based on blind rechecking and panel testing results, PPM-DOTS site laboratories showed good agreement with the reference laboratory. Physical space and qualities of AFB reagents would be areas of intervention to sustain the quality of sputum smear microscopy. Therefore, regular external quality assessment and provision of basic laboratory supplies for TB diagnosis would be the way forward to improve the quality of sputum smear microscopy services in PPM-DOTS laboratories. PMID:25849516

  5. Pharmacodynamic Modeling of Bacillary Elimination Rates and Detection of Bacterial Lipid Bodies in Sputum to Predict and Understand Outcomes in Treatment of Pulmonary Tuberculosis

    PubMed Central

    Sloan, Derek J.; Mwandumba, Henry C.; Garton, Natalie J.; Khoo, Saye H.; Butterworth, Anthony E.; Allain, Theresa J.; Heyderman, Robert S.; Corbett, Elizabeth L.; Barer, Mike R.; Davies, Geraint R.

    2015-01-01

    Background. Antibiotic-tolerant bacterial persistence prevents treatment shortening in drug-susceptible tuberculosis, and accumulation of intracellular lipid bodies has been proposed to identify a persister phenotype of Mycobacterium tuberculosis cells. In Malawi, we modeled bacillary elimination rates (BERs) from sputum cultures and calculated the percentage of lipid body–positive acid-fast bacilli (%LB + AFB) on sputum smears. We assessed whether these putative measurements of persistence predict unfavorable outcomes (treatment failure/relapse). Methods. Adults with pulmonary tuberculosis received standard 6-month therapy. Sputum samples were collected during the first 8 weeks for serial sputum colony counting (SSCC) on agar and time-to positivity (TTP) measurement in mycobacterial growth indicator tubes. BERs were extracted from nonlinear and linear mixed-effects models, respectively, fitted to these datasets. The %LB + AFB counts were assessed by fluorescence microscopy. Patients were followed until 1 year posttreatment. Individual BERs and %LB + AFB counts were related to final outcomes. Results. One hundred and thirty-three patients (56% HIV coinfected) participated, and 15 unfavorable outcomes were reported. These were inversely associated with faster sterilization phase bacillary elimination from the SSCC model (odds ratio [OR], 0.39; 95% confidence interval [CI], .22–.70) and a faster BER from the TTP model (OR, 0.71; 95% CI, .55–.94). Higher %LB + AFB counts on day 21–28 were recorded in patients who suffered unfavorable final outcomes compared with those who achieved stable cure (P = .008). Conclusions. Modeling BERs predicts final outcome, and high %LB + AFB counts 3–4 weeks into therapy may identify a persister bacterial phenotype. These methods deserve further evaluation as surrogate endpoints for clinical trials. PMID:25778753

  6. Entamoeba gingivalis in sputum smears.

    PubMed

    Dao, A H

    1985-01-01

    Entamoeba gingivalis is a common parasite of the human buccal cavity whose rare appearance in Papanicolaou-stained sputum smears may be missed. Two such cases are described, including the morphologic features of this ameba. The trophozoites were seen to phagocytize leukocytes as well as red blood cells, in distinction to E. histiolytica, which phagocytizes only red blood cells and also can cause pulmonary abscesses. The concomitant finding of Actinomyces sp. organisms in one patient reinforces the possible symbiotic relationship between the two organisms, as has been suggested for their appearance in other extraoral sites, such as the female genital tract. PMID:3861055

  7. Novel hypertonic saline-sodium hydroxide (HS-SH) method for decontamination and concentration of sputum samples for Mycobacterium tuberculosis microscopy and culture.

    PubMed

    Ganoza, Christian A; Ricaldi, Jessica N; Chauca, José; Rojas, Gabriel; Munayco, César; Agapito, Juan; Palomino, Juan Carlos; Guerra, Humberto

    2008-09-01

    This study evaluated a new decontamination and concentration (DC) method for sputum microscopy and culture. Sputum samples from patients with suspected pulmonary tuberculosis (TB) (n=106) were tested using the proposed hypertonic saline-sodium hydroxide (HS-SH) DC method, the recommended N-acetyl-L-cysteine-sodium citrate-sodium hydroxide (NALC-NaOH) DC method and unconcentrated direct smear (Ziehl-Neelsen) techniques for the presence of mycobacteria using Löwenstein-Jensen culture and light microscopy. Of 94 valid specimens, 21 (22.3%) were positive in culture and were further characterized as Mycobacterium tuberculosis. The sensitivity for acid-fast bacilli (AFB) smears was increased from 28.6% using the direct method to 71.4% (HS-SH) and 66.7% (NALC-NaOH) using DC methods. Both concentration techniques were highly comparable for culture (kappa=0.794) and smear (kappa=0.631) for AFB. Thus the proposed HS-SH DC method improved the sensitivity of AFB microscopy compared with a routine unconcentrated direct smear; its performance was comparable to that of the NALC-NaOH DC method for AFB smears and culture, but it was methodologically simpler and less expensive, making it a promising candidate for evaluation by national TB control programmes in developing countries.

  8. Natural attenuation study at Columbus AFB MS

    SciTech Connect

    Stauffer, T.; Libelo, E.; MacIntyre, W.; Boggs, J.

    1995-12-31

    In order to study the geochemical and biochemical processes which contribute to natural attenuation of hydrocarbons in ground water systems, a subsurface residual NAPL hydrocarbon mixture was emplaced in the well characterized and highly instrumented heterogeneous aquifer at the Columbus AFB, MS groundwater test site. 1,147 kg of NAPL composed of decane, naphthalene, p-xylene, ethylbenzene, toluene, benzene and 2 Kg of KBr tracer was mixed with 30 m{sup 3} of local aquifer material to create a 16% residual phase and emplaced below the water table on November 23rd, 1995. Natural hydraulic gradients are now dissolving the hydrocarbons and transporting the dissolved hydrocarbon and bromide plume. Background sampling of groundwater and aquifer solids was done prior to source emplacement to characterize the site geochemistry and anaerobic and aerobic microbiology. The aquifer was initially oxygenated with DO levels ranging from 0.5 to 6.9 mg/L and generally < 3.5, NO{sub 3}-N ranged from 0.02--0.3 mg/L. Sulfate concentrations ranged from 0.0 to 8.6 mg/L. Dissolved Fe{sup 2+} ranged up to 5.0 mg/L. Observed natural attenuation rates will be correlated with microbial and geochemical changes in the aquifer. These correlations will provide a basis for understanding and implementing natural attenuation as a remedial action for hydrocarbons.

  9. Mishap Investigation Team (MIT) - Barksdale AFB, Louisiana

    NASA Technical Reports Server (NTRS)

    Stepaniak, Philip

    2005-01-01

    The Shuttle Program is organized to support a Shuttle mishap using the resources of the MIT. The afternoon of Feb. 1, 2003, the MIT deployed to Barksdale AFB. This location became the investigative center and interim storage location for crewmembers received from the Lufkin Disaster Field Office (DFO). Working under the leadership of the MIT Lead, the medical team executed a short-term plan that included search, recovery, and identification including coordination with the Armed Forces Institute of Pathology Temporary operations was set up at Barksdale Air Force Base for two weeks. During this time, coordination with the DFO field recovery teams, AFIP personnel, and the crew surgeons was on going. In addition, the crewmember families and NASA management were updated daily. The medical team also dealt with public reports and questions concerning biological and chemical hazards, which were coordinated with SPACEHAB, Inc., Kennedy Space Center (KSC) Medical Operations and the Johnson Space Center (JSC) Space Medicine office. After operations at Barksdale were concluded the medical team transitioned back to Houston and a long-term search, recovery and identification plan was developed.

  10. Similar activation state of neutrophils in sputum of asthma patients irrespective of sputum eosinophilia.

    PubMed

    Tak, T; Hilvering, B; Tesselaar, K; Koenderman, L

    2015-11-01

    Inflammatory phenotypes of asthma are associated with differences in disease characteristics. It is unknown whether these inflammatory phenotypes are reflected by the activation status of neutrophils in blood and sputum. We obtained peripheral blood and induced sputum from 21 asthma patients and stratified our samples based on sputum eosinophilia resulting in two groups (>3% eosinophils: n = 13, <3%: n = 8). Eosinophils and neutrophils from blood and sputum were analysed for expression of activation and degranulation markers by flow cytometry. Data were analysed by both classical, non-parametric statistics and a multi-dimensional approach, using principal component analysis (PCA). Patients with sputum eosinophilia were characterized by increased asthma control questionnaire (ACQ) scores and blood eosinophil counts. Both sputum neutrophils and eosinophils displayed an activated and degranulated phenotype compared to cells obtained from blood. Specifically, degranulation of all granule types was detected in sputum cells, combined with an increased expression of the activation markers (activated) Mac-1 (CD11b), programmed death ligand 1 (PD-L1) (CD274) and a decreased expression of CD62L. CD69 expression was only increased on sputum eosinophils. Surface marker expression of neutrophils was similar in the presence or absence of eosinophilia, either by single or multi-dimensional analysis. Sputum neutrophils were highly activated and degranulated irrespective of sputum eosinophilia. Therefore, we conclude that differences in granulocyte activation in sputum and/or blood are not associated with clinical differences in the two groups of asthma patients. The finding of PD-L1 expression on sputum granulocytes suggests an immunomodulatory role of these cells in the tissue.

  11. The Arabidopsis Auxin Receptor F-Box Proteins AFB4 and AFB5 Are Required for Response to the Synthetic Auxin Picloram.

    PubMed

    Prigge, Michael J; Greenham, Kathleen; Zhang, Yi; Santner, Aaron; Castillejo, Cristina; Mutka, Andrew M; O'Malley, Ronan C; Ecker, Joseph R; Kunkel, Barbara N; Estelle, Mark

    2016-01-01

    The plant hormone auxin is perceived by a family of F-box proteins called the TIR1/AFBs. Phylogenetic studies reveal that these proteins fall into four clades in flowering plants called TIR1, AFB2, AFB4, and AFB6. Genetic studies indicate that members of the TIR1 and AFB2 groups act as positive regulators of auxin signaling by promoting the degradation of the Aux/IAA transcriptional repressors. In this report, we demonstrate that both AFB4 and AFB5 also function as auxin receptors based on in vitro assays. We also provide genetic evidence that AFB4 and AFB5 are targets of the picloram family of auxinic herbicides in addition to indole-3-acetic acid. In contrast to previous studies we find that null afb4 alleles do not exhibit obvious defects in seedling morphology or auxin hypersensitivity. We conclude that AFB4 and AFB5 act in a similar fashion to other members of the family but exhibit a distinct auxin specificity. PMID:26976444

  12. The Arabidopsis Auxin Receptor F-Box Proteins AFB4 and AFB5 Are Required for Response to the Synthetic Auxin Picloram

    PubMed Central

    Prigge, Michael J.; Greenham, Kathleen; Zhang, Yi; Santner, Aaron; Castillejo, Cristina; Mutka, Andrew M.; O’Malley, Ronan C.; Ecker, Joseph R.; Kunkel, Barbara N.; Estelle, Mark

    2016-01-01

    The plant hormone auxin is perceived by a family of F-box proteins called the TIR1/AFBs. Phylogenetic studies reveal that these proteins fall into four clades in flowering plants called TIR1, AFB2, AFB4, and AFB6. Genetic studies indicate that members of the TIR1 and AFB2 groups act as positive regulators of auxin signaling by promoting the degradation of the Aux/IAA transcriptional repressors. In this report, we demonstrate that both AFB4 and AFB5 also function as auxin receptors based on in vitro assays. We also provide genetic evidence that AFB4 and AFB5 are targets of the picloram family of auxinic herbicides in addition to indole-3-acetic acid. In contrast to previous studies we find that null afb4 alleles do not exhibit obvious defects in seedling morphology or auxin hypersensitivity. We conclude that AFB4 and AFB5 act in a similar fashion to other members of the family but exhibit a distinct auxin specificity. PMID:26976444

  13. Characteristics of endobronchial tuberculosis patients with negative sputum acid-fast bacillus

    PubMed Central

    Yıldız, Pınar

    2013-01-01

    Objective Endobronchial tuberculosis (EBTB) is defined as a tuberculous infection of the tracheobronchial tree with microbial and histopathological evidence, with or without parenchymal involvement. In this study, clinical, radiological and bronchoscopic characteristics of cases diagnosed to have EBTB were evaluated. Methods Sixteen patients with at least three negative sputum examinations for acid-fast bacillus (AFB) and diagnosed as having EBTB on the histopathological examination of bronchoscopically obtained specimens showing granulomatous structures with caseation necrosis and/or positive AFB-culture on the microbiological examination of bronchoscopically obtained specimens were included in our study. Age, sex, symptoms, tuberculin skin test (TST), microbiological examination results and radiological findings were recorded. Bronchoscopical lesions were classified according to Chung classification. Results EBTB was found to be more common in females. Most common symptoms were cough (100%), sputum (75%), weight loss (62.5%), hemoptisis (37.5%), chest pain (25%) and dyspnea (12.5%). Radiological examination findings revealed consolidations/infiltrations (87.5%), nodular lesions (37.5%), cavitary lesions (25%), unilateral (43.7%) or bilateral hilar widening (31.2%) and atelectasia (25%). Middle lob syndrome was seen in three cases. Most common lesions observed bronchoscopically were active caseous lesions, granular lesions, edematous hyperemic lesions, tumorous lesions, fibrostenotic lesions respectively. In all cases “granulomatous inflammation showing caseation” was shown in the histopathological examination of biopsy specimens. Conclusions EBTB can cause various radiological and bronchoscopical findings. In most of the cases distinct response is seen to antituberculous treatment. Bronchial stenosis is an important complication. Treatment should be given as soon as possible to avoid it. PMID:24409353

  14. The effect of green tea extract supplementation on sputum smear conversion and weight changes in pulmonary TB patients: A randomized controlled trial

    PubMed Central

    Honarvar, Mohammad Reza; Eghtesadi, Shahryar; Gill, Pooria; Jazayeri, Shima; Vakili, Mohammad Ali; Shamsardekani, Mohammad Reza; Abbasi, Abdollah

    2016-01-01

    Background: Acceleration in sputum smear conversion helps faster improvement and decreased probability of the transfer of TB. In this study, we aimed to investigate the effect of green tea extract supplementation on sputum smear conversion and weight changes in smear positive pulmonary TB patients in Iran. Methods: In this double blind clinical study, TB patients were divided into intervention, (n=43) receiving 500 mg green tea extract (GTE), and control groups (n=40) receiving placebo for two months, using balanced randomization. Random allocation and allocation concealment were observed. Height and weight were measured at the beginning, and two and six months post-treatment. Evaluations were performed on three slides, using the ZiehlNeelsen method. Independent and paired t test, McNemar’s, Wilcoxon, Kaplan-Meier, Cox regression model and Log-Rank test were utilized. Statistical significance was set at p<0.05. This trial was registered under IRCT201212232602N11. Results: The interventional changes and the interactive effect of intervention on weight were not significant (p>0.05). In terms of shortening the duration of conversion, the case to control proportion showed a significant difference (p=0.032). Based on the Cox regression model, the hazard ratio of the relative risk of delay in sputum smear conversion was 3.7 (p=0.002) in the higher microbial load group compared to the placebo group and 0.54 (95% CI: 0.31-0.94) in the intervention compared to the placebo group. Conclusion: GTE decreases the risk of delay in sputum smear conversion, but has no effect on weight gain. Moreover, it may be used as an adjuvant therapy for faster rehabilitation for pulmonary TB patients. PMID:27493925

  15. High Genotypic Discordance of Concurrent Mycobacterium tuberculosis Isolates from Sputum and Blood of HIV-Infected Individuals

    PubMed Central

    Ssengooba, Willy; Cobelens, Frank G.; Nakiyingi, Lydia; Mboowa, Gerald; Armstrong, Derek T.; Manabe, Yukari C.; Joloba, Moses L.; de Jong, Bouke C.

    2015-01-01

    Background Among HIV-infected individuals with CD4 less than 200 cells/mm3, tuberculosis often has an atypical presentation, is more likely to be disseminated and is diagnostically challenging. We sought to understand the genotypic discordance of concurrent sputum and blood M. tuberculosis (MTB) isolates from HIV-infected individuals. Methods From a prospective diagnostic accuracy study with 182 HIV-infected culture-positive TB adults, isolates were obtained from 51 of 66 participants who were MTB culture-positive by both sputum and blood. Isolates were subjected to susceptibility testing to 1st line drugs, spoligotyping and 24 locus- MIRU-VNTR. Results The median age of the participants was 31 (IQR; 27–38) years and 51% were male. The median CD4 count was 29 (IQR; 10–84) cells/mm3 with 20% taking ART; 8.0% were previously treated for TB, and 63% were AFB smear-negative. The isolates belonged to two of the main global MTB-lineages; East-African-Indian (L3) 17 (16.7%) and Euro-American (L4) 85 (83.3%). We identified 26 (51.0%) participants with discordant MTB-genotypes between sputum and blood, including two patients with evidence of mixed infection in either compartment. Having discordant MTB-genotypes was not predicted by the MTB-lineage in either blood or sputum, CD4 cell count, or any other clinical characteristic. Conclusions There is a high genotypic discordance among M. tuberculosis concurrently isolated from sputum and blood of HIV-infected individuals. These findings suggest that infection with more than one strain of M. tuberculosis occurs in at least half of patients with advanced HIV infection. PMID:26176604

  16. Improved polyacrylamide-based artificial sputum with formalin-fixed tubercle bacilli for training of tuberculosis microscopists.

    PubMed

    Yamada, Hiroyuki; Mitarai, Satoshi; Wahyunitisari, Manik Rento; Mertaniasih, Ni Made; Sugamoto, Tetsuhiro; Chikamatsu, Kinuyo; Aono, Akio; Matsumoto, Hiroko; Fujiki, Akiko

    2011-10-01

    Sputum smear microscopy is an easy, inexpensive, and rapid method for detecting tubercle bacilli when there are more than 10,000 bacilli/ml in the original sputum. Furthermore, because the microscopic method provides not only quantitative, but also qualitative information, such as the shape of bacilli, it has remained significant. We have previously developed and reported panel test slides made from polyacrylamide-based artificial sputum (PBAS) mixed with both cultured THP-1 cells and nonpathogenic mycobacteria. In this paper, we report an improved preparation method for PBAS for panel test slides that provides a simplified method and enhanced availability with high consistency in each grade and in which only negative PBAS is prepared from polyacrylamide and cultured THP-1 cells and mixed with graded formalin-fixed Mycobacterium tuberculosis solution (FFTBS) containing oral flora and Pseudomonas aeruginosa on the slides. In the smears prepared using this improved method, the numbers (average ± standard deviation [SD]) of acid-fast bacilli (AFB) in 300 fields (2- by 3-cm smear) in eight smears of each grade ranged from 5 to 9 (6.4 ± 1.4), from 59 to 88 (74.6 ± 10.0), from 503 to 912 (705.0 ± 145.7), and from 1,819 to 3,256 (2133.3 ± 478.0) in ±, +, ++, and +++ smears, respectively. In addition, this preparation method provided high similarity to the microscopic appearance of bacilli and background seen in the actual patient sputum, with high feasibility. These results revealed that our new PBAS had high authenticity in the appearance and consistency in each grade, which could make it valuable as a reliable artificial sputum for the training of microscopists.

  17. Improved Polyacrylamide-Based Artificial Sputum with Formalin-Fixed Tubercle Bacilli for Training of Tuberculosis Microscopists▿†

    PubMed Central

    Yamada, Hiroyuki; Mitarai, Satoshi; Wahyunitisari, Manik Rento; Mertaniasih, Ni Made; Sugamoto, Tetsuhiro; Chikamatsu, Kinuyo; Aono, Akio; Matsumoto, Hiroko; Fujiki, Akiko

    2011-01-01

    Sputum smear microscopy is an easy, inexpensive, and rapid method for detecting tubercle bacilli when there are more than 10,000 bacilli/ml in the original sputum. Furthermore, because the microscopic method provides not only quantitative, but also qualitative information, such as the shape of bacilli, it has remained significant. We have previously developed and reported panel test slides made from polyacrylamide-based artificial sputum (PBAS) mixed with both cultured THP-1 cells and nonpathogenic mycobacteria. In this paper, we report an improved preparation method for PBAS for panel test slides that provides a simplified method and enhanced availability with high consistency in each grade and in which only negative PBAS is prepared from polyacrylamide and cultured THP-1 cells and mixed with graded formalin-fixed Mycobacterium tuberculosis solution (FFTBS) containing oral flora and Pseudomonas aeruginosa on the slides. In the smears prepared using this improved method, the numbers (average ± standard deviation [SD]) of acid-fast bacilli (AFB) in 300 fields (2- by 3-cm smear) in eight smears of each grade ranged from 5 to 9 (6.4 ± 1.4), from 59 to 88 (74.6 ± 10.0), from 503 to 912 (705.0 ± 145.7), and from 1,819 to 3,256 (2133.3 ± 478.0) in ±, +, ++, and +++ smears, respectively. In addition, this preparation method provided high similarity to the microscopic appearance of bacilli and background seen in the actual patient sputum, with high feasibility. These results revealed that our new PBAS had high authenticity in the appearance and consistency in each grade, which could make it valuable as a reliable artificial sputum for the training of microscopists. PMID:21813720

  18. Improved polyacrylamide-based artificial sputum with formalin-fixed tubercle bacilli for training of tuberculosis microscopists.

    PubMed

    Yamada, Hiroyuki; Mitarai, Satoshi; Wahyunitisari, Manik Rento; Mertaniasih, Ni Made; Sugamoto, Tetsuhiro; Chikamatsu, Kinuyo; Aono, Akio; Matsumoto, Hiroko; Fujiki, Akiko

    2011-10-01

    Sputum smear microscopy is an easy, inexpensive, and rapid method for detecting tubercle bacilli when there are more than 10,000 bacilli/ml in the original sputum. Furthermore, because the microscopic method provides not only quantitative, but also qualitative information, such as the shape of bacilli, it has remained significant. We have previously developed and reported panel test slides made from polyacrylamide-based artificial sputum (PBAS) mixed with both cultured THP-1 cells and nonpathogenic mycobacteria. In this paper, we report an improved preparation method for PBAS for panel test slides that provides a simplified method and enhanced availability with high consistency in each grade and in which only negative PBAS is prepared from polyacrylamide and cultured THP-1 cells and mixed with graded formalin-fixed Mycobacterium tuberculosis solution (FFTBS) containing oral flora and Pseudomonas aeruginosa on the slides. In the smears prepared using this improved method, the numbers (average ± standard deviation [SD]) of acid-fast bacilli (AFB) in 300 fields (2- by 3-cm smear) in eight smears of each grade ranged from 5 to 9 (6.4 ± 1.4), from 59 to 88 (74.6 ± 10.0), from 503 to 912 (705.0 ± 145.7), and from 1,819 to 3,256 (2133.3 ± 478.0) in ±, +, ++, and +++ smears, respectively. In addition, this preparation method provided high similarity to the microscopic appearance of bacilli and background seen in the actual patient sputum, with high feasibility. These results revealed that our new PBAS had high authenticity in the appearance and consistency in each grade, which could make it valuable as a reliable artificial sputum for the training of microscopists. PMID:21813720

  19. AFB/open cycle gas turbine conceptual design study

    NASA Technical Reports Server (NTRS)

    Dickinson, T. W.; Tashjian, R.

    1983-01-01

    Applications of coal fired atmospheric fluidized bed gas turbine systems in industrial cogeneration are identified. Based on site-specific conceptual designs, the potential benefits of the AFB/gas turbine system were compared with an atmospheric fluidized design steam boiler/steam turbine system. The application of these cogeneration systems at four industrial plant sites is reviewed. A performance and benefit analysis was made along with a study of the representativeness of the sites both in regard to their own industry and compared to industry as a whole. A site was selected for the conceptual design, which included detailed site definition, AFB/gas turbine and AFB/steam turbine cogeneration system designs, detailed cost estimates, and comparative performance and benefit analysis. Market and benefit analyses identified the potential market penetration for the cogeneration technologies and quantified the potential benefits.

  20. Characterization of Afb, a novel bifunctional protein in Streptococcus agalactiae

    PubMed Central

    Dehbashi, Sanaz; Pourmand, Mohammad Reza; Mashhadi, Rahil

    2016-01-01

    Background and Objectives: Streptococcus agalactiae is the leading cause of bacterial sepsis and meningitis in newborns and results in pneumonia and bacteremia in adults. A number of S. agalactiae components are involved in colonization of target cells. Destruction of peptidoglycan and division of covalently linked daughter cells is mediated by autolysins. In this study, autolytic activity and plasma binding ability of AFb novel recombinant protein of S. agalactiae was investigated. Materials and Methods: The gbs1805 gene was cloned and expressed. E. coli strains DH5α and BL21 were used as cloning and expression hosts, respectively. After purification, antigenicity and binding ability to plasma proteins of the recombinant protein was evaluated. Results: AFb, the 18KDa protein was purified successfully. The insoluble mature protein revealed the ability to bind to fibrinogen and fibronectin. This insoluble mature protein revealed that it has the ability to bind to fibrinogen and fibronectin plasma proteins. Furthermore, in silico analysis demonstrated the AFb has an autolytic activity. Conclusions: AFb is a novel protein capable of binding to fibrinogen and fibronectin. This findings lay a ground work for further investigation of the role of the bacteria in adhesion and colonization to the host. PMID:27092228

  1. SOLVENT CHEMICAL INVENTORY OF THE NEWARK AFB BUILDING 4 FACILITY

    EPA Science Inventory

    A solvent chemical inventory was performed at Newark AFB to develop a chemical usage profile of the Building 4 facility, and identify primary sources of solvent emissions. This survey was performed in two parts. Part 1 consisted of a chemical inventory survey which defined solv...

  2. Standardization of a TaqMan-Based Real-Time PCR for the Detection of Mycobacterium tuberculosis-Complex in Human Sputum

    PubMed Central

    Barletta, Francesca; Vandelannoote, Koen; Collantes, Jimena; Evans, Carlton A.; Arévalo, Jorge; Rigouts, Leen

    2014-01-01

    Real-time polymerase chain reaction (qPCR) was optimized for detecting Mycobacterium tuberculosis in sputum. Sputum was collected from patients (N = 112) with suspected pulmonary tuberculosis, tested by smear microscopy, decontaminated, and split into equal aliquots that were cultured in Löwenstein-Jensen medium and tested by qPCR for the small mobile genetic element IS6110. The human ERV3 sequence was used as an internal control. 3 of 112 (3%) qPCR failed. For the remaining 109 samples, qPCR diagnosed tuberculosis in 79 of 84 patients with culture-proven tuberculosis, and sensitivity was greater than microscopy (94% versus 76%, respectively, P < 0.05). The qPCR sensitivity was similar (P = 0.9) for smear-positive (94%, 60 of 64) and smear-negative (95%, 19 of 20) samples. The qPCR was negative for 24 of 25 of the sputa with negative microscopy and culture (diagnostic specificity 96%). The qPCR had 99.5% sensitivity and specificity for 211 quality control samples including 84 non-tuberculosis mycobacteria. The qPCR cost ∼5US$ per sample and provided same-day results compared with 2–6 weeks for culture. PMID:25114009

  3. Standardization of a TaqMan-based real-time PCR for the detection of Mycobacterium tuberculosis-complex in human sputum.

    PubMed

    Barletta, Francesca; Vandelannoote, Koen; Collantes, Jimena; Evans, Carlton A; Arévalo, Jorge; Rigouts, Leen

    2014-10-01

    Real-time polymerase chain reaction (qPCR) was optimized for detecting Mycobacterium tuberculosis in sputum. Sputum was collected from patients (N = 112) with suspected pulmonary tuberculosis, tested by smear microscopy, decontaminated, and split into equal aliquots that were cultured in Löwenstein-Jensen medium and tested by qPCR for the small mobile genetic element IS6110. The human ERV3 sequence was used as an internal control. 3 of 112 (3%) qPCR failed. For the remaining 109 samples, qPCR diagnosed tuberculosis in 79 of 84 patients with culture-proven tuberculosis, and sensitivity was greater than microscopy (94% versus 76%, respectively, P < 0.05). The qPCR sensitivity was similar (P = 0.9) for smear-positive (94%, 60 of 64) and smear-negative (95%, 19 of 20) samples. The qPCR was negative for 24 of 25 of the sputa with negative microscopy and culture (diagnostic specificity 96%). The qPCR had 99.5% sensitivity and specificity for 211 quality control samples including 84 non-tuberculosis mycobacteria. The qPCR cost ∼5US$ per sample and provided same-day results compared with 2-6 weeks for culture.

  4. 76 FR 55553 - Amendment of Class D Airspace; Eglin AFB, FL

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-09-08

    ... Airspace in the Eglin Air Force Base (AFB), FL airspace area. The Destin Non-Directional Beacon (NDB) has... airport. This action also corrects the geographic coordinates of Eglin AFB, Duke Field, and Hulbert Field... D airspace in the Eglin AFB, FL area (76 FR 38580) Docket No. FAA-2011-0087. Subsequent...

  5. The Epidemiology and Geographic Distribution of Nontuberculous Mycobacteria Clinical Isolates from Sputum Samples in the Eastern Region of China

    PubMed Central

    Song, Honghuan; Li, Guoli; Liu, Qiao; Li, Yan; Zhu, Limei; Martinez, Leonardo; Lu, Wei

    2015-01-01

    Background Nontuberculous mycobacteria (NTM) have been reported to be increasing worldwide and its geographic distribution differs by region. The aim of this study was to describe the epidemiology and distribution of NTM in the eastern part of China. Methods Sputum samples were collected from 30 surveillance sites for tuberculosis drug resistance test from May 1, 2008 to December 31, 2008. Identification was performed using a biochemical test, multiplex PCR and GenoType Mycobacterium CM/AS assay. Results A total of 1779 smear positive clinical isolates were obtained, of which 60 (3.37%) were NTM. Five species/complex of NTM were identified; M. intracellulare was the predominated species (68.33%), followed by M. abscessus-M. immunogenum (13.33%), Mycobacterium spec. (10.00%), M. Kansasii (6.67%) and M. peregrinum-M. alvei-M. septicum (1.67%). Conclusion M. intracellulare was the main species of NTM in the eastern part of China and clinical physicians should pay more attention to NTM induced pulmonary disease. PMID:25775117

  6. A Rapid and Sensitive Diagnostic Screening Assay for Detection of Mycobacteria Including Mycobacterium tuberculosis Directly from Sputum without Extraction.

    PubMed

    Cross, Lisa Jane; Anscombe, Catherine; McHugh, Timothy D; Abubakar, Ibrahim; Shorten, Robert John; Thorne, Nicola; Arnold, Cath

    2015-01-01

    We report a novel approach utilising a real-time PCR screening assay targeting a 53 bp tandemly repeated element present at various loci within the Mycobacterium tuberculosis (Mtb) genome. Positive samples were identified within a discriminatory melting curve range of 90-94°C, with results obtained in under one hour directly from decontaminated sputum samples without extraction. A panel of 89 smear-positive sputa were used for analytical validation of the assay with 100% concordance, with sensitivity matching that of culture. Cross reactivity was detected within a narrow range of mycobacteria other than tuberculosis (MOTT) (five sputa, three in silico), with the highest sensitivity within M. avium complex (MAC). A year-long head to head evaluation of the test with the GeneXpert platform was carried out with 104 consecutive samples at the Royal Free Hospital, UK. Receiver operating characteristics (ROC) analysis of the data revealed that the two tests are approximately equivalent in sensitivity, with the area under the curve being 0.85 and 0.80 for the GeneXpert and our assay, respectively, indicating that the test would be a cost effective screen prior to GeneXpert testing.

  7. Relationship between sputum inflammatory markers and osmotic airway hyperresponsiveness during induction of sputum in asthmatic patients.

    PubMed Central

    Jang, A. S.; Choi, I. S.

    2001-01-01

    Hypertonic saline aerosols are being used increasingly for bronchial provocation testing and induction of sputum. The aims of this study were to assess the response to challenge with 3% hypertonic saline administered via a ultrasonic nebulizer in patients with asthma, and to evaluate relationship between % fall of FEV1 during induction of sputum (osmotic airway hyperresponsiveness; osmotic AHR) and biochemical markers of induced sputum. We investigated changes in FEV1 in response to inhaling ultrasonically nebulized 3% saline in 25 patients with asthma and 10 control subjects. FEV1 was measured before, during, and after induction of sputum. We used fluoroimmunoassay to detect eosinophil cationic protein (ECP), immunohistochemical staining to detect EG2+ (secretory form of ECP) eosinophils, and a sandwich ELISA to detect interleukin (IL)-5. Protein concentration was determined by using bicinchoninic acid protein assay reagent. Asthmatics, compared with controls, had significantly higher osmotic AHR. Moderate to severe asthmatics had significantly higher osmotic AHR compared to mild asthmatics. Osmotic AHR was significantly correlated with the proportion of eosinophils, the levels of ECP, EG2+ eosinophils, IL-5, and proteins. These data suggest that osmotic AHR is closely related to the clinical status and biochemical markers of sputum supernatant in asthmatic patients. PMID:11511785

  8. External quality assessment of AFB smear microscopy performances and its associated factors in selected private health facilities in Addis Ababa, Ethiopia.

    PubMed

    Mosissa, Lemi; Kebede, Abebaw; Mindaye, Tedla; Getahun, Muluwork; Tulu, Sisay; Desta, Kassu

    2016-01-01

    Tuberculosis (TB) is still a public health problem in sub Saharan African countries. In resource-limited settings, TB diagnosis relies on sputum smear microscopy, with low and variable sensitivities, especially in paucibacillary pediatric and HIV-associated TB patients. Tuberculosis microscopy centers have several weaknesses like overworking, insufficiently trained personnel, inconsistent reagent supplies, and poorly maintained equipments; thus, there is a critical need for investments in laboratory infrastructure, capacity building, and quality assurance schemes. The performance of TB microscopy centers in the private health facilities in Addis Ababa is not known so far. The main objective of the study was to assess laboratory performance of acid fast bacilli (AFB) smear microscopy and its associated factors in selected private health facilities in Addis Ababa, Ethiopia. A cross-sectional study was conducted in 33 selected private health facilities of Addis Ababa, Ethiopia comprising 7 hospitals, 2 NGO health centers, 23 higher clinics and 1 diagnostic laboratory that provide AFB smear microscopy services. The study was conducted from January to April 2014. A total of 283 stained sputum smears were randomly collected from participant laboratories for blinded rechecking, 320 panel slides were sent to 32 microscopy centers to evaluate their performance on AFB reading, staining and reporting. Checklists were used to assess quality issues of laboratories. Data were captured, cleaned, and analyzed using SPSS version 16.0; χ(2) tests, kappa statistics were used for comparison purpose. P value < 0.05 considered statistically significant. Among the 32 participant laboratories, 2-scored 100%, 15 scored 80-95% & the remaining 15 scored 50-75% for overall proficiency test performance. There were 10 (3.15%) major errors and 121 (37.8%) minor errors. The sensitivity, specificity, PPV and NPV of panel reading by microscopy centers were 89%, 96%, 96%, and 90% respectively. Out

  9. External quality assessment of AFB smear microscopy performances and its associated factors in selected private health facilities in Addis Ababa, Ethiopia.

    PubMed

    Mosissa, Lemi; Kebede, Abebaw; Mindaye, Tedla; Getahun, Muluwork; Tulu, Sisay; Desta, Kassu

    2016-01-01

    Tuberculosis (TB) is still a public health problem in sub Saharan African countries. In resource-limited settings, TB diagnosis relies on sputum smear microscopy, with low and variable sensitivities, especially in paucibacillary pediatric and HIV-associated TB patients. Tuberculosis microscopy centers have several weaknesses like overworking, insufficiently trained personnel, inconsistent reagent supplies, and poorly maintained equipments; thus, there is a critical need for investments in laboratory infrastructure, capacity building, and quality assurance schemes. The performance of TB microscopy centers in the private health facilities in Addis Ababa is not known so far. The main objective of the study was to assess laboratory performance of acid fast bacilli (AFB) smear microscopy and its associated factors in selected private health facilities in Addis Ababa, Ethiopia. A cross-sectional study was conducted in 33 selected private health facilities of Addis Ababa, Ethiopia comprising 7 hospitals, 2 NGO health centers, 23 higher clinics and 1 diagnostic laboratory that provide AFB smear microscopy services. The study was conducted from January to April 2014. A total of 283 stained sputum smears were randomly collected from participant laboratories for blinded rechecking, 320 panel slides were sent to 32 microscopy centers to evaluate their performance on AFB reading, staining and reporting. Checklists were used to assess quality issues of laboratories. Data were captured, cleaned, and analyzed using SPSS version 16.0; χ(2) tests, kappa statistics were used for comparison purpose. P value < 0.05 considered statistically significant. Among the 32 participant laboratories, 2-scored 100%, 15 scored 80-95% & the remaining 15 scored 50-75% for overall proficiency test performance. There were 10 (3.15%) major errors and 121 (37.8%) minor errors. The sensitivity, specificity, PPV and NPV of panel reading by microscopy centers were 89%, 96%, 96%, and 90% respectively. Out

  10. External quality assessment of AFB smear microscopy performances and its associated factors in selected private health facilities in Addis Ababa, Ethiopia

    PubMed Central

    Mosissa, Lemi; Kebede, Abebaw; Mindaye, Tedla; Getahun, Muluwork; Tulu, Sisay; Desta, Kassu

    2016-01-01

    Tuberculosis (TB) is still a public health problem in sub Saharan African countries. In resource-limited settings, TB diagnosis relies on sputum smear microscopy, with low and variable sensitivities, especially in paucibacillary pediatric and HIV-associated TB patients. Tuberculosis microscopy centers have several weaknesses like overworking, insufficiently trained personnel, inconsistent reagent supplies, and poorly maintained equipments; thus, there is a critical need for investments in laboratory infrastructure, capacity building, and quality assurance schemes. The performance of TB microscopy centers in the private health facilities in Addis Ababa is not known so far. The main objective of the study was to assess laboratory performance of acid fast bacilli (AFB) smear microscopy and its associated factors in selected private health facilities in Addis Ababa, Ethiopia. A cross-sectional study was conducted in 33 selected private health facilities of Addis Ababa, Ethiopia comprising 7 hospitals, 2 NGO health centers, 23 higher clinics and 1 diagnostic laboratory that provide AFB smear microscopy services. The study was conducted from January to April 2014. A total of 283 stained sputum smears were randomly collected from participant laboratories for blinded rechecking, 320 panel slides were sent to 32 microscopy centers to evaluate their performance on AFB reading, staining and reporting. Checklists were used to assess quality issues of laboratories. Data were captured, cleaned, and analyzed using SPSS version 16.0; χ2 tests, kappa statistics were used for comparison purpose. P value < 0.05 considered statistically significant. Among the 32 participant laboratories, 2-scored 100%, 15 scored 80-95% & the remaining 15 scored 50-75% for overall proficiency test performance. There were 10 (3.15%) major errors and 121 (37.8%) minor errors. The sensitivity, specificity, PPV and NPV of panel reading by microscopy centers were 89%, 96%, 96%, and 90% respectively. Out of

  11. Building Energy Audit Report, for Hickam AFB, HI

    SciTech Connect

    Chvala, William D.; De La Rosa, Marcus I.; Brown, Daryl R.; Dixon, Douglas R.

    2010-09-30

    A building energy assessment was performed by a team of engineers from Pacific Northwest National Laboratory (PNNL) under contract to the Department of Energy/Federal Energy Management program (FEMP). The effort used the Facility Energy Decision System (FEDS) model to determine how energy is consumed at Hickam AFB, identify the most cost-effective energy retrofit measures, and calculate the potential energy and cost savings. This documents reports the results of that assessment.

  12. Procedures for Collection of Induced Sputum Specimens From Children

    PubMed Central

    Grant, Lindsay R.; Murdoch, David R.; O’Brien, Katherine L.; Scott, J. Anthony

    2012-01-01

    In most settings, sputum is not routinely collected for microbiological diagnosis from children with lower respiratory disease. To evaluate whether it is feasible and diagnostically useful to collect sputum in the Pneumonia Etiology Research for Child Health (PERCH) study, we reviewed the literature on induced sputum procedures. Protocols for induced sputum in children were collated from published reports and experts on respiratory disease and reviewed by an external advisory group for recommendation in the PERCH study. The advisory group compared 6 protocols: 4 followed a nebulization technique using hypertonic saline, and 2 followed a chest or abdomen massage technique. Grading systems for specimen quality were evaluated. Collecting sputum from children with lower respiratory tract illness is feasible and is performed around the world. An external advisory group recommended that sputum be collected from children hospitalized with severe and very severe pneumonia who participate in the PERCH study provided no contraindications exist. PERCH selected the nebulization technique using hypertonic saline. PMID:22403228

  13. Acoustofluidic Transfer of Inflammatory Cells from Human Sputum Samples.

    PubMed

    Li, Sixing; Ren, Liqiang; Huang, Po-Hsun; Yao, Xianglan; Cuento, Rosemarie A; McCoy, J Philip; Cameron, Craig E; Levine, Stewart J; Huang, Tony Jun

    2016-06-01

    For sputum analysis, the transfer of inflammatory cells from liquefied sputum samples to a culture medium or buffer solution is a critical step because it removes the inflammatory cells from the presence of residual dithiothreitol (DTT), a reagent that reduces cell viability and interferes with further sputum analyses. In this work, we report an acoustofluidic platform for transferring inflammatory cells using standing surface acoustic waves (SSAW). In particular, we exploit the acoustic radiation force generated from a SSAW field to actively transfer inflammatory cells from a solution containing residual DTT to a buffer solution. The viability and integrity of the inflammatory cells are maintained during the acoustofluidic-based cell transfer process. Our acoustofluidic technique removes residual DTT generated in sputum liquefaction and facilitates immunophenotyping of major inflammatory cells from sputum samples. It enables cell transfer in a continuous flow, which aids the development of an automated, integrated system for on-chip sputum processing and analysis. PMID:27183317

  14. Griffiss AFB integrated resource assessment. Volume 2, Electric baseline detail

    SciTech Connect

    Dixon, D.R.; Armstrong, P.R.; Keller, J.M.

    1993-02-01

    The US Air Force Air Combat Command has tasked the Pacific Northwest Laboratory (PNL) as the lead laboratory supporting the US Department of Energy (DOE) Federal Energy Management Program`s (FEMP) mission to identify, evaluate, and assist in acquiring all cost-effective energy projects at Griffiss Air Force Base (AFB). This is a model program PNL is designing for federal customers served by the Niagara Mohawk Power Company (Niagara Mohawk). It will (1) identify and evaluate all electric cost-effective energy projects; (2) develop a schedule at each installation for project acquisition considering project type, size, timing, and capital requirements, as well as energy and dollar savings; and (3) secure 100% of the financing required to implement electric energy efficiency projects from Niagara Mohawk and have Niagara Mohawk procure the necessary contractors to perform detailed audits and install the technologies. This report documents the assessment of baseline energy use at one of Niagara Mohawk`s primary federal facilities, Griffiss AFB, an Air Combat Command facility located near Rome, New York. It is a companion report to Volume 1, the Executive Summary, and Volume 3, the Electric Resource Assessment. The analysis examines the characteristics of electric, gas, oil, propane, coal, and purchased thermal capacity use for fiscal year (FY) 1990. The results include energy-use intensities for the facilities at Griffiss AFB by building type and electric energy end use. A complete electric energy consumption reconciliation is presented that accounts for the distribution of all major electric energy uses and losses among buildings, utilities, and central systems.

  15. Endeavour lands at Edwards AFB, ending mission STS-100

    NASA Technical Reports Server (NTRS)

    2001-01-01

    NASA/Edwards AFB, Calif. -- With its drag chute deployed, Endeavour lands on runway 22 at Edwards Air Force Base, Calif., at 12:10:42 p.m. EDT after a mission of 11 days, 12 hours, 54 minutes to the International Space Station on mission STS-100. The orbiter and its crew of seven logged about 4.9 million statute miles in 186 orbits. Due to unfavorable weather conditions, landing at KSC was waved off. The landing marked the third consecutive landing at EAFB.

  16. Spectra analysis of coating antigen: A possible explanation for difference in anti-AFB1 polyclonal antibody sensitivity

    NASA Astrophysics Data System (ADS)

    Ye, Yang; Liu, Aiping; Wang, Xiaohong; Chen, Fusheng

    2016-10-01

    For the detection of small hapten molecules, indirect competitive enzyme-linked immunosorbent assay (icELISA) is a preferred method. However, diverse coating antigen might bring different antiserum titer and sensitivity for the identical antiserum. In the present study, four AFB1-protein (aflatoxin B1-carrier protein) conjugates were prepared by activated ester method (AFB1O-BSA/AFB1O-OVA) and mannich method (AFB1-cBSA/AFB1-cOVA), and then applied as coating antigen for titer and sensitivity detection of the identical antiserum obtained from rabbit immunized by AFB1-KLH. Afterwards, the ultraviolet-visible, fluorescence and far-ultraviolet circular dichroism (far-UV CD) spectra were recorded for understanding the difference in titer and sensitivity obtained. Results revealed that AFB1O-BSA/AFB1O-OVA showed a strong intrinsic fluorescence band centered at 450 nm that originated from the emission of AFB1, which differed from AFB1-cBSA/AFB1-cOVA, while the decrease of α-helical and increase of β-sheet in AFB1-cBSA was the most remarkable. This indicated that the better sensitivity obtained by using AFB1O-BSA as coating antigen might be caused by its extended structure, because such structure affect the binding between AFB1 and antibody. The study might offer structural information for understanding the titer and sensitivity difference caused by coating antigen.

  17. Giant African Pouched Rats as Detectors of Tuberculosis in Human Sputum: Comparison of Two Techniques for Sputum Presentation

    ERIC Educational Resources Information Center

    Wray, Alisha M.; Mahoney, Amanda; Weetjens, Bart J.; Cox, Christophe; Jubitana, Maureen; Kazwala, Rudovic; Mfinanga, Godfrey S.; Durgin, Amy; Poling, Alan

    2013-01-01

    Previous studies have shown that pouched rats can detect the presence of "Mycobacterium tuberculosis," which causes tuberculosis, in human sputum samples obtained from clinical facilities. Although pouched rats evaluate sputum samples quickly, preparing the samples is relatively slow. The present study evaluated whether the rats can detect…

  18. Identification and validation of nebulized aerosol devices for sputum induction

    PubMed Central

    Davidson, Warren J; Dennis, John; The, Stephanie; Litoski, Belinda; Pieron, Cora; Leigh, Richard

    2014-01-01

    Induced sputum cell count measurement has proven reliability for evaluating airway inflammation in patients with asthma and other airway diseases. Although the use of nebulizer devices for sputum induction is commonplace, they are generally labelled as single-patient devices by the manufacturer and, therefore, cannot be used for multiple patients in large clinical sputum induction programs due to infect ion-control requirements. Accordingly, this study investigated the aerosol characteristics of alternative devices that could be used in such programs. BACKGROUND: Induced sputum cell counts are a noninvasive and reliable method for evaluating the presence, type and degree of airway inflammation in patients with asthma. Currently, standard nebulizer devices used for sputum induction in multiple patients are labelled as single-patient devices by the manufacturer, which conflicts with infection prevention and control requirements. As such, these devices cannot feasibly be used in a clinical sputum induction program. Therefore, there is a need to identify alternative nebulizer devices that are either disposable or labelled for multi-patient use. OBJECTIVE: To apply validated rigorous, scientific testing methods to identify and validate commercially available nebulizer devices appropriate for use in a clinical sputum induction program. METHODS: Measurement of nebulized aerosol output and size for the selected nebulizer designs followed robust International Organization for Standardization methods. Sputum induction using two of these nebulizers was successfully performed on 10 healthy adult subjects. The cytotechnologist performing sputum cell counts was blinded to the type of nebulizer used. RESULTS: The studied nebulizers had variable aerosol outputs. The AeroNeb Solo (Aerogen, Ireland), Omron NE-U17 (Omron, Japan) and EASYneb II (Flaem Nuova, Italy) systems were found to have similar measurements of aerosol size. There was no significant difference in induced sputum

  19. Treatment Default amongst Patients with Tuberculosis in Urban Morocco: Predicting and Explaining Default and Post-Default Sputum Smear and Drug Susceptibility Results

    PubMed Central

    Ghali, Iraqi; Kizub, Darya; Billioux, Alexander C.; Bennani, Kenza; Bourkadi, Jamal Eddine; Benmamoun, Abderrahmane; Lahlou, Ouafae; Aouad, Rajae El; Dooley, Kelly E.

    2014-01-01

    Setting Public tuberculosis (TB) clinics in urban Morocco. Objective Explore risk factors for TB treatment default and develop a prediction tool. Assess consequences of default, specifically risk for transmission or development of drug resistance. Design Case-control study comparing patients who defaulted from TB treatment and patients who completed it using quantitative methods and open-ended questions. Results were interpreted in light of health professionals’ perspectives from a parallel study. A predictive model and simple tool to identify patients at high risk of default were developed. Sputum from cases with pulmonary TB was collected for smear and drug susceptibility testing. Results 91 cases and 186 controls enrolled. Independent risk factors for default included current smoking, retreatment, work interference with adherence, daily directly observed therapy, side effects, quick symptom resolution, and not knowing one’s treatment duration. Age >50 years, never smoking, and having friends who knew one’s diagnosis were protective. A simple scoring tool incorporating these factors was 82.4% sensitive and 87.6% specific for predicting default in this population. Clinicians and patients described additional contributors to default and suggested locally-relevant intervention targets. Among 89 cases with pulmonary TB, 71% had sputum that was smear positive for TB. Drug resistance was rare. Conclusion The causes of default from TB treatment were explored through synthesis of qualitative and quantitative data from patients and health professionals. A scoring tool with high sensitivity and specificity to predict default was developed. Prospective evaluation of this tool coupled with targeted interventions based on our findings is warranted. Of note, the risk of TB transmission from patients who default treatment to others is likely to be high. The commonly-feared risk of drug resistance, though, may be low; a larger study is required to confirm these findings

  20. Endeavour lands at Edwards AFB, ending mission STS-100

    NASA Technical Reports Server (NTRS)

    2001-01-01

    NASA/Edwards AFB, Calif. -- After landing at Edwards Air Force Base, Calif., the STS-100 crew poses for a photograph in front of orbiter Endeavour, which successfully launched them to the International Space Station and returned them to Earth. They are (left to right) Mission Specialists John Phillips, Umberto Guidoni and Chris Hadfield; Pilot Jeffrey Ashby; Commander Kent Rominger; and Mission Specialists Yuri Lonchakov and Scott Parazynski. Guidoni is with the European Space Agency, Hadfield with the Canadian Space Agency and Lonchakov with the Russian Aviation and Space Agency. The orbiter and crew logged about 4.9 million statute miles in 186 orbits. Due to unfavorable weather conditions, landing at KSC was waved off. The landing marked the third consecutive landing at EAFB.

  1. McClellan AFB Management Action Plan (MAP) submittal

    SciTech Connect

    Not Available

    1992-12-23

    This Management Action Plan ('Action Plan' or 'MAP') contains a status summary of the McClellan Air Force Base (McAFB) environmental restoration and compliance programs and presents a comprehensive strategy for implementing response actions necessary to protect human health and the environment. This strategy integrates activities under both the Installation Restoration Program (IRP) and the Environmental Compliance Program (ECP). This Action Plan is a dynamic document that will be updated on a regular basis using the change-a-page looseleaf binder concept for day-to-day revisions along with a subsection at the end of each chapter to highlight any modifications or innovations since the previous major annual review/update.

  2. Shemya AFB, Alaska 1992 IRP field investigation report

    SciTech Connect

    Not Available

    1993-02-01

    The US Air Force is currently investigating 22 sites on Shemya Air Force Base (AFB) to determine if past spill and disposal activities have caused environmental damage. These investigations are being carried out under the Air Force's Installation Restoration Program (IRP). As a part of the IRP program, field investigations were performed in 1992 to obtain the information needed to assess what future actions willneed to be carried out at each site. The island's drinking water supply was also investigated. Activities completed at 10 selected sites during the 1992 field investigation included surface sampling to determine the lateral extent of contamination, subsurface sampling to determine the vertical extent of contamination, and the installation of well points and monitoring wells to determine the direction of groundwater flow and if the groundwater has been affected by a site. In addition, geophysical surveys were performed at most sites to identify site boundaries and check for the presence of buried metal to be avoided during drilling activities.

  3. Exophiala Pneumonia Presenting with a Cough Productive of Black Sputum

    PubMed Central

    Cohen, Yehuda Z.; Stead, Wendy

    2015-01-01

    Exophiala species are black, yeast-like molds that can cause subcutaneous cysts as well as disseminated disease. Isolated pneumonia due to Exophiala species is extremely uncommon. We report a case of isolated Exophiala pneumonia in a patient with bronchiectasis who presented with worsening dyspnea and a cough productive of black sputum. The production of black sputum, known as melanoptysis, is an uncommon physical finding with a limited differential diagnosis. To our knowledge, this is the first reported case of Exophiala pneumonia presenting with a cough productive of black sputum. PMID:26075119

  4. Exophiala Pneumonia Presenting with a Cough Productive of Black Sputum.

    PubMed

    Cohen, Yehuda Z; Stead, Wendy

    2015-01-01

    Exophiala species are black, yeast-like molds that can cause subcutaneous cysts as well as disseminated disease. Isolated pneumonia due to Exophiala species is extremely uncommon. We report a case of isolated Exophiala pneumonia in a patient with bronchiectasis who presented with worsening dyspnea and a cough productive of black sputum. The production of black sputum, known as melanoptysis, is an uncommon physical finding with a limited differential diagnosis. To our knowledge, this is the first reported case of Exophiala pneumonia presenting with a cough productive of black sputum. PMID:26075119

  5. Aerospace medicine at Brooks AFB, TX: hail and farewell.

    PubMed

    Nunneley, Sarah A; Webb, James T

    2011-05-01

    With the impending termination of USAF operations at Brooks Air Force Base (AFB) in San Antonio, TX, it is time to consider its historic role in Aerospace Medicine. The base was established in 1917 as a flight training center for the U.S. Army Air Service and in 1926 became home to its School of Aviation Medicine. The school moved to San Antonio's Randolph Field in 1931, but in 1959 it returned to Brooks where it occupied new facilities to support its role as a national center for U.S. Air Force aerospace medicine, including teaching, clinical medicine, and research. The mission was then expanded to encompass support of U.S. military and civilian space programs. With the abrupt termination of the military space program in 1969, research at Brooks focused on clinical aviation medicine and support of advanced military aircraft while continuing close cooperation with NASA in support of orbital spaceflight and the journey to the Moon. Reorganization in the 1990s assigned all research functions at Brooks to the Human Systems Division and its successors, leaving to USAFSAM the missions related to clinical work and teaching. In 2002 the USAF and the city of San Antonio implemented shared operation of Brooks as a "City-Base" in the hope of deflecting threatened closure. Nevertheless, under continuing pressure to consolidate military facilities in the United States, the 2005 Base Closure and Realignment Commission ordered Brooks closed by 2011, with its aerospace medicine functions relocated to new facilities at Wright-Patterson AFB in Dayton, OH.

  6. IRON-BINDING AND STORAGE PROTEINS IN SPUTUM

    EPA Science Inventory

    Induced sputum (IS) and bronchoalveolar lavage (BAL) sample different lung compartments, with IS obtaining secretions from the surfaces of the bronchial airways and BAL sampling secretions from the alveolar airspaces. Deposition of iron-containing particulate matter occurs prefer...

  7. Isolation of Staphylococcus aureus from sputum in cystic fibrosis.

    PubMed

    Sparham, P D; Lobban, D I; Speller, D C

    1978-10-01

    The success in the isolation of Staphylococcus aureus of different methods of sputum processing was investigated in 60 specimens collected from 14 patients with cystic fibrosis during a seven-month period. Fifty specimens (83%) from 11 patients yielded Staph. aureus by one or more methods. Direct plating of purulent portions of sputum on to media designed for general use in respiratory infections gave unsatisfactory results (35% yield of Staph. aureus). Some increase in isolations was obtained with preliminary liquefaction of sputum; but the best results were given by the addition of a medium selective for staphylococci (mannitol salt agar, BBL) or by initial sonication of sputum (each 83% yield). Seven of the 11 strains of Staph. aureus were thymidine-dependent and otherwise atypical in laboratory characteristics; these were isolated from patients who had received co-trimoxazole. PMID:101553

  8. Induced sputum evaluation in restorers and conservators of cultural heritage.

    PubMed

    Kanceljak-Macan, Božica; Trošić, Ivančica; Varnai, Veda Marija; Pavičić, Ivan; Macan, Jelena

    2012-01-01

    The objective of this study was to examine induced sputum (IS) cells profile from restorers/conservators of cultural heritage (restorers) with no lower airway symptoms and normal ventilatory lung function. The study involved 22 restorers and 48 control workers. Medical interview, physical examination, spirometry, skin prick testing to inhalatory allergens, and IS collection were performed. Compared with control workers, restorers demonstrated higher percentage of neutrophils (34% vs 15.5%; p = .004). This pattern was found only in male workers. They had almost 9 times greater chance to have increased proportion of sputum neutrophils than male controls (odds ratio [OR] 8.98, 95% confidence interval [CI] 1.98-40.7). The mobilization of eosinophils or macrophages into the airways was not established regardless of workers sex. Additional gender difference in sputum cells distribution was found for occupationally unexposed subjects, with higher proportion of sputum neutrophils in women.

  9. Isolation of oxidase-negative Pseudomonas aeruginosa from sputum culture.

    PubMed Central

    Hampton, K D; Wasilauskas, B L

    1979-01-01

    Two isolates of Pseudomonas aeruginosa lacking characteristic indophenol oxidase were recovered from a sputum specimen. A discussion of the characteristic biochemical tests and antibiograms along with a possible explanation for this phenomenon is presented. PMID:225349

  10. Thioredoxin liquefies and decreases the viscoelasticity of cystic fibrosis sputum.

    PubMed

    Rancourt, Raymond C; Tai, Shusheng; King, Malcolm; Heltshe, Sonya L; Penvari, Churee; Accurso, Frank J; White, Carl W

    2004-05-01

    The persistent and viscous nature of airway secretions in cystic fibrosis (CF) disease leads to airway obstruction, opportunistic infection, and deterioration of lung function. Thioredoxin (Trx) is a protein disulfide reductase that catalyzes numerous thiol-dependent cellular reductive processes. To determine whether Trx can alter the rheological properties of mucus, sputum obtained from CF patients was treated with TRX and its reducing system (0.1 microM thioredoxin reductase + 2 mM NADPH), and liquid phase-gel phase ratio (percent liquid phase) was assessed by compaction assay. Exposure to low Trx concentrations (1 microM) caused significant increases in the percentage of liquid phase of sputum. Maximal increases in percent liquid phase occurred with 30 microM Trx. Additional measurements revealed that sputum liquefaction by the Trx reducing system is dependent on NADPH concentration. The relative potency of the Trx reducing system also was compared with other disulfide-reducing agents. In contrast with Trx, glutathione and N-acetylcysteine were ineffective in liquefying sputum when used at concentrations <1 mM. Sputum viscoelasticity, measured by magnetic microrheometry, also was diminished significantly following 20-min treatment with 3, 10, or 30 microM Trx. Similarly, this reduction in viscoelasticty also was dependent on NADPH concentration. Further investigation has indicated that Trx treatment increases the solubility of high-molecular-weight glycoproteins and causes redistribution of extracellular DNA into the liquid phase of sputum. Recognizing that mucins are the major gel-forming glycoproteins in mucus, we suggest that Trx alters sputum rheology by enzymatic reduction of glycoprotein polymers present in sputum. PMID:14695120

  11. Bioremediation of a Large Chlorinated Solvent Plume, Dover AFB, DE

    SciTech Connect

    Bloom, Aleisa C

    2015-01-01

    Bioremediation of a Large Chlorinated Solvent Plume, Dover AFB, DE Aleisa Bloom, (Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA) Robert Lyon (bob.lyon@aecom.com), Laurie Stenberg, and Holly Brown (AECOM, Germantown, Maryland, USA) ABSTRACT: Past disposal practices at Dover Air Force Base (AFB), Delaware, created a large solvent plume called Area 6 (about 1 mile long, 2,000 feet wide, and 345 acres). The main contaminants are PCE, TCE, and their degradation products. The remedy is in-situ accelerated anaerobic bioremediation (AAB). AAB started in 2006 and is focusing on source areas and downgradient plume cores. Direct-push injections occurred in source areas where contamination is typically between 5 and 20 feet below ground surface. Lower concentration dissolved-phased contamination is present downgradient at 35 and 50 feet below ground surface. Here, permanent injection/extraction wells installed in transects perpendicular to the flow of groundwater are used to apply AAB. The AAB substrate is a mix of sodium lactate, emulsified vegetable oil, and nutrients. After eight years, dissolved contaminant mass within the main 80-acre treatment area has been reduced by over 98 percent. This successful application of AAB has stopped the flux of contaminants to the more distal portions of the plume. While more time is needed for effects to be seen in the distal plume, AAB injections will soon cease, and the remedy will transition to natural attenuation. INTRODUCTION Oak Ridge National Laboratory Environmental Science Division (ORNL) and AECOM (formerly URS Corporation) have successfully implemented in situ accelerated anaerobic bioremediation (AAB) to remediate chlorinated solvent contamination in a large, multi-sourced groundwater plume at Dover Air Force Base (AFB). AAB has resulted in significant reductions of dissolved phase chlorinated solvent concentrations. This plume, called Area 6, was originally over 1 mile in length and over 2,000 feet wide (Figure 1

  12. AFB as a discovery tool for Z' bosons at the LHC

    NASA Astrophysics Data System (ADS)

    Accomando, E.; Belyaev, A.; Fiaschi, J.; Mimasu, K.; Moretti, S.; Shepherd-Themistocleous, C.

    2016-07-01

    The Forward-Backward Asymmetry (AFB) in Z' physics is commonly only perceived as the observable which possibly allows one to interpret a Z' signal by distinguishing different models of such (heavy) spin-1 bosons. In this article, we examine the potential of AFB in setting bounds on or even discovering a Z' at the Large Hadron Collider (LHC) and show that it might be a powerful tool for this purpose. We analyze two different scenarios: Z' s with a narrow and wide width, respectively. We find that in both cases AFB can complement the cross section in accessing Z' signals.

  13. AFB1-induced mutagenesis of the gpt gene in AS52 cells

    PubMed Central

    Wattanawaraporn, Roongtiwa; Kim, Min Young; Adams, Jillian; Trudel, Laura J.; Woo, Leslie L.; Croy, Robert G.; Essigmann, John M.; Wogan, Gerald N.

    2013-01-01

    Aflatoxin B1 (AFB1) is a potent mutagen and an important risk factor for hepatocellular carcinoma (HCC) in humans. Transgenic mouse strains and cells in culture have been used to detect different types of mutations caused by AFB1 and investigate the molecular determinants of their location and frequency. The AFB1 mutational spectrum in the gpt gene was markedly different in AS52 cells compared to the liver in gpt delta B6C3F1 transgenic mice. The results demonstrate the importance of metabolism, chromosomal location, transcription and selection conditions on mutational spectra. PMID:22733615

  14. Shemya AFB, Alaska 1992 IRP field investigation report

    SciTech Connect

    Not Available

    1993-02-01

    The US Air Force is currently investigating 22 sites on Shemya Air Force Base (AFB) to determine if past spill and disposal activities have caused environmental damage. These investigations are being carried out under the Air Force's Installation Restoration Program (IRP). Field investigations were performed in 1992 to obtain the information needed to assess what future actions will need to be carried out at each site. The island's drinking water supply was also investigated. Activities completed at 10 selected sites included surface sampling to determine the lateral extent of contamination, subsurface sampling to determine the vertical extent of contamination, and the installation of well points and monitoring wells to determine the direction of groundwater flow and if the groundwater has been affected by a site. Geophysical surveys were performed at most sites to identify site boundaries and check for the presence of buried metal to be avoided during drilling activities. This report, appendices B, C, and D contains information on the following: geophysical contour maps and profile plots; human health risk assessment; and ecological risk assessment.

  15. Shemya AFB, Alaska 1992 IRP field investigation report

    SciTech Connect

    Not Available

    1993-02-01

    The US Air Force is currently investigating 22 sites on Shemya Air Force Base (AFB) to determine if past spill and disposal activities have caused environmental damage. These investigations are being carried out under the Air Force's Installation Restoration Program (IRP). Field investigations were performed in 1992 to obtain the information needed to assess what future actions will need to be carried out at each site. The island's drinking water supply was also investigated. Activities completed at 10 selected sites during the 1992 field investigation included surface sampling to determine the lateral extent of contamination, subsurface sampling to determine the vertical extent of contamination, and the installation of well points and monitoring wells to determine the direction of groundwater flow and if the groundwater has been affected by a site. In addition, geophysical surveys were performed at most sites to identify site boundaries and check for the presence of buried metal to be avoided during drilling activities. This report, appendix A, contains the analytical results.

  16. Shemya AFB, Alaska 1992 IRP field investigation report

    SciTech Connect

    Not Available

    1993-02-01

    The US Air Force is currently investigating 22 sites on Shemya Air Force Base (AFB) to determine if past spill and disposal activities have caused environmental damage. These investigations are being carried out under the Air Force's Installation Restoration Program (IRP). Field investigations were performed in 1992 to obtain the information needed to assess what future actions will need to be carried out at each site. The island's drinking water supply was also investigated. Activities completed at 10 selected sites included surface sampling to determine the lateral extent of contamination, subsurface sampling to determine the vertical extent of contamination, and the installation of well points and monitoring wells to determine the direction of groundwater flow and if the groundwater has been affected by a site. In addition, geophysical surveys were performed at most sites to identify site boundaries and check for the presence of buried metal, to be avoided during drilling activities. This report contains appendices E and F with information on the following: soil boring logs, and data validation of samples analyzed.

  17. Dover AFB characterization/hazardous-waste management survey, Dover AFB, Delaware. Final report, 25 February-31 May 1986

    SciTech Connect

    Binovi, R.D.; Ng, E.K.; Slavich, F.E.

    1986-07-01

    The USAFOEHL conducted an on-site wastewater-characterization survey at Dover AFB, DE from 25 Feb to 7 Mar 86. The survey was designed to establish pretreatment requirements by determining quantities and concentrations of pollutants expected from the industrial operations or develop and evaluate alternate solutions to decrease the discharge of wastewater contaminants. Effluents from industrial and domestic wastewater were analyzed. Kent County DE has imposed stringent pretreatment standards by changing the sampling location from a point where industrial wastewater is combined with considerably more domestic wastewater to a point where only industrial wastewater combines. Effluent limitations of cadmium were exceeded during each of the seven days sampling. Sources of cadmium were found to be metal fabrication, corrosion control, and vehicle-maintenance operations. Recommendations: 1) Clean lift station sumps. 2) Change method of stripping. 3) Install pretreatment operation to remove chromium and cadium from the wastewater prior to discharge into the sanitary sewer. 4) Perform periodic EP Toxicity testing on neutralized battery acid. 5) Repipe vats in building 719 to provide piping dedicated to each vat. 6) Negotiate with a solvent-recovery representative to provide a system for a trial period.

  18. [Usefulness of sputum Gram staining in community-acquired pneumonia].

    PubMed

    Sato, Tadashi; Aoshima, Masahiro; Ohmagari, Norio; Tada, Hiroshi; Chohnabayashi, Naohiko

    2002-07-01

    To evaluate the usefulness of sputum gram staining in community-acquired pneumonia (CAP), we reviewed 144 cases requiring hospitalization in the last 4 years. The sensitivity was 75.5%, specificity 68.2%, positive predictive value 74.1%, negative predictive value 69.8%, positive likelihood ratio 2.37, negative likelihood ratio 0.36 and accuracy 72.2% in 97 cases. Both sputum gram staining and culture were performed. Concerning bacterial pneumonia (65 cases), we compared the Gram staining group (n = 33), which received initial antibiotic treatment, based on sputum gram staining with the Empiric group (n = 32) that received antibiotics empirically. The success rates of the initial antibiotic treatment were 87.9% vs. 78.1% (P = 0.473); mean hospitalization periods were 9.67 vs. 11.75 days (P = 0.053); and periods of intravenous therapy were 6.73 vs. 7.91 days (P = 0.044), respectively. As for initial treatment, penicillins were used in the Gram staining group more frequently (P < 0.01). We conclude that sputum gram staining is useful for the shortening of the treatment period and the appropriate selection of initial antibiotics in bacterial pneumonia. We believe, therefore, that sputum gram staining is indispensable as a diagnostic tool CAP.

  19. Alveolar-cell carcinoma: a problem in sputum cytodiagnosis.

    PubMed Central

    Spriggs, A I; Cole, M; Dunnill, M S

    1982-01-01

    Cytology and histology are correlated in a series of 22 cases chosen to illustrate the differential diagnosis between clusters of benign bronchial or bronchiolar cells seen in sputum, and those of alveolar cell carcinoma or adenocarcinoma with alveolar spread. Alveolar-cell carcinoma is characterised by clusters of small epithelial cells in spherical or irregular formations, none showing enough polarity to distinguish a smooth or palisaded surface. The appearances are most distinctive if vacuolation is absent. The diagnosis cannot, however, be confidently made in all cases from morphological features of cells in sputum. Images PMID:6294147

  20. Microstructural alterations of sputum in cystic fibrosis lung disease

    PubMed Central

    Duncan, Gregg A.; Jung, James; Joseph, Andrea; Thaxton, Abigail L.; West, Natalie E.; Boyle, Michael P.; Hanes, Justin

    2016-01-01

    The stasis of mucus secretions in the lungs of cystic fibrosis (CF) patients leads to recurrent infections and pulmonary exacerbations, resulting in decreased survival. Prior studies have assessed the biochemical and biophysical features of airway mucus in individuals with CF. However, these measurements are unable to probe mucus structure on microscopic length scales relevant to key players in the progression of CF-related lung disease, namely, viruses, bacteria, and neutrophils. In this study, we quantitatively determined sputum microstructure based on the diffusion of muco-inert nanoparticle probes in CF sputum and found that a reduction in sputum mesh pore size is characteristic of CF patients with reduced lung function, as indicated by measured FEV1. We also discovered that the effect of ex vivo treatment of CF sputum with rhDNase I (Pulmozyme) on microstructure is dependent upon the time interval between the most recent inhaled rhDNase I treatment and the sample collection. Microstructure of mucus may serve as a marker for the extent of CF lung disease and as a parameter for assessing the effectiveness of mucus-altering agents. PMID:27812540

  1. Characterization of sputum biomarkers for asthma–COPD overlap syndrome

    PubMed Central

    Gao, Jing; Iwamoto, Hiroshi; Koskela, Jukka; Alenius, Harri; Hattori, Noboru; Kohno, Nobuoki; Laitinen, Tarja; Mazur, Witold; Pulkkinen, Ville

    2016-01-01

    Asthma–COPD overlap syndrome (ACOS) is a commonly encountered chronic airway disease. However, ACOS is still a consensus-based clinical phenotype and the underlying inflammatory mechanisms are inadequately characterized. To clarify the inflammatory mediatypical for ACOS, five biomarkers, namely interleukin (IL)-13, myeloperoxidase (MPO), neutrophil gelatinase-associated lipocalin (NGAL), chitinase-like protein (YKL-40), and IL-6, were selected. This study hypothesized that sputum biomarkers relevant for airway inflammation in asthma (IL-13), COPD (MPO, NGAL), or in both asthma and COPD (YKL-40, IL-6) could be used to differentiate ACOS from COPD and asthma. The aim of this study was to characterize the inflammatory profile and improve the recognition of ACOS. Induced sputum levels of IL-13, MPO, NGAL, YKL-40, and IL-6 were measured by enzyme-linked immunosorbent assay/Luminex assay in a Finnish discovery cohort (n=90) of nonsmokers, smokers, and patients with asthma, COPD, and ACOS and validated in a Japanese cohort (n=135). The classification accuracy of potential biomarkers was compared with area under the receiver operating characteristic curves. Only sputum NGAL levels could differentiate ACOS from asthma (P<0.001 and P<0.001) and COPD (P<0.05 and P=0.002) in the discovery and replication cohorts, respectively. Sputum NGAL levels were independently correlated with the percentage of pre-bronchodilator forced expiratory volume in 1 second predicted in multivariate analysis in the discovery and replication cohorts (P=0.001 and P=0.002, respectively). In conclusion, sputum biomarkers reflecting both airway inflammation and remodeling of the tissue show potential in differentiation between asthma, COPD, and ACOS. PMID:27757028

  2. Reliability of a rapid hematology stain for sputum cytology*

    PubMed Central

    Gonçalves, Jéssica; Pizzichini, Emilio; Pizzichini, Marcia Margaret Menezes; Steidle, Leila John Marques; Rocha, Cristiane Cinara; Ferreira, Samira Cardoso; Zimmermann, Célia Tânia

    2014-01-01

    Objective: To determine the reliability of a rapid hematology stain for the cytological analysis of induced sputum samples. Methods: This was a cross-sectional study comparing the standard technique (May-Grünwald-Giemsa stain) with a rapid hematology stain (Diff-Quik). Of the 50 subjects included in the study, 21 had asthma, 19 had COPD, and 10 were healthy (controls). From the induced sputum samples collected, we prepared four slides: two were stained with May-Grünwald-Giemsa, and two were stained with Diff-Quik. The slides were read independently by two trained researchers blinded to the identification of the slides. The reliability for cell counting using the two techniques was evaluated by determining the intraclass correlation coefficients (ICCs) for intraobserver and interobserver agreement. Agreement in the identification of neutrophilic and eosinophilic sputum between the observers and between the stains was evaluated with kappa statistics. Results: In our comparison of the two staining techniques, the ICCs indicated almost perfect interobserver agreement for neutrophil, eosinophil, and macrophage counts (ICC: 0.98-1.00), as well as substantial agreement for lymphocyte counts (ICC: 0.76-0.83). Intraobserver agreement was almost perfect for neutrophil, eosinophil, and macrophage counts (ICC: 0.96-0.99), whereas it was moderate to substantial for lymphocyte counts (ICC = 0.65 and 0.75 for the two observers, respectively). Interobserver agreement for the identification of eosinophilic and neutrophilic sputum using the two techniques ranged from substantial to almost perfect (kappa range: 0.91-1.00). Conclusions: The use of Diff-Quik can be considered a reliable alternative for the processing of sputum samples. PMID:25029648

  3. Diagnostic value of microscopic examination of Gram-stained sputum and sputum cultures in patients with bacteremic pneumococcal pneumonia.

    PubMed

    Musher, Daniel M; Montoya, Roberto; Wanahita, Anna

    2004-07-15

    Clinicians continue to question the usefulness of microscopic examination of Gram-stained sputum specimens ("Gram staining") and sputum culture for diagnosis of pneumonia. We analyzed the sensitivity of these techniques in 105 patients with pneumococcal pneumonia proven by blood culture. Gram staining revealed gram-positive cocci in pairs and chains, and culture yielded pneumococci in only 31% and 44% of all cases, respectively. However, sputum specimens were never submitted for examination in 31 cases; in 16 others, the specimen was inadequate and a culture was not done. Excluding these cases, the sensitivities of Gram staining and culture were 57% and 79%, respectively. If patients receiving antibiotics for >24 h had been excluded, Gram staining would have suggested pneumococci in 63%, and culture results would have been positive in 86%. Sensitivity increased in inverse proportion to the duration of antibiotic therapy (P<.05). Microscopic examination of sputum samples before antibiotics were administered and performance of culture within 24 h of receipt of such treatment yielded the correct diagnosis in >80% of cases of pneumococcal pneumonia.

  4. Analysis of Lung Flute–collected Sputum for Lung Cancer Diagnosis

    PubMed Central

    Su, Jian; Anjuman, Nigar; Guarnera, Maria A; Zhang, Howard; Stass, Sanford A; Jiang, Feng

    2015-01-01

    Molecular analysis of sputum can help diagnose lung cancer. We have demonstrated that Lung Flute can be used to collect sputum from individuals who cannot spontaneously expectorate sputum. The objective of this study is to further evaluate the performance of the Lung Flute by comparing the characteristics of parallel samples collected with and without the Lung Flute and the usefulness for diagnosis of lung cancer. Fifty-six early-stage lung cancer patients (40 current smokers and 16 former smokers) and 73 cancer-free individuals (52 current smokers and 21 former smokers) were instructed to spontaneously cough and use Lung Flute for sputum sampling. Sputum cytology and polymerase chain reaction analysis of three miRNAs (miRs-21, 31, and 210) were performed in the specimens. All 92 current smokers and 11 (28.7%) of 37 former smokers spontaneously expectorated sputum and also produced sputum when using the Lung Flute. Twenty-seven former smokers (70.3%) who could not spontaneously expectorate sputum, however, were able to produce sputum when using the Lung Flute. The specimens were of low respiratory origin without contamination from other sources, eg, saliva. There was no difference of sputum volume and cell populations, diagnostic efficiency of cytology, and analysis of the miRNAs in the specimens collected by the two approaches. Analysis of the sputum miRNAs produced 83.93% sensitivity and 87.67% specificity for identifying lung cancer. Therefore, sputum collected by the Lung Flute has comparable features as spontaneously expectorated sputum. Using the Lung Flute enables former smokers who cannot spontaneously expectorate to provide adequate sputum to improve sputum collection for lung cancer diagnosis. PMID:26309391

  5. Analysis of Lung Flute-collected Sputum for Lung Cancer Diagnosis.

    PubMed

    Su, Jian; Anjuman, Nigar; Guarnera, Maria A; Zhang, Howard; Stass, Sanford A; Jiang, Feng

    2015-01-01

    Molecular analysis of sputum can help diagnose lung cancer. We have demonstrated that Lung Flute can be used to collect sputum from individuals who cannot spontaneously expectorate sputum. The objective of this study is to further evaluate the performance of the Lung Flute by comparing the characteristics of parallel samples collected with and without the Lung Flute and the usefulness for diagnosis of lung cancer. Fifty-six early-stage lung cancer patients (40 current smokers and 16 former smokers) and 73 cancer-free individuals (52 current smokers and 21 former smokers) were instructed to spontaneously cough and use Lung Flute for sputum sampling. Sputum cytology and polymerase chain reaction analysis of three miRNAs (miRs-21, 31, and 210) were performed in the specimens. All 92 current smokers and 11 (28.7%) of 37 former smokers spontaneously expectorated sputum and also produced sputum when using the Lung Flute. Twenty-seven former smokers (70.3%) who could not spontaneously expectorate sputum, however, were able to produce sputum when using the Lung Flute. The specimens were of low respiratory origin without contamination from other sources, eg, saliva. There was no difference of sputum volume and cell populations, diagnostic efficiency of cytology, and analysis of the miRNAs in the specimens collected by the two approaches. Analysis of the sputum miRNAs produced 83.93% sensitivity and 87.67% specificity for identifying lung cancer. Therefore, sputum collected by the Lung Flute has comparable features as spontaneously expectorated sputum. Using the Lung Flute enables former smokers who cannot spontaneously expectorate to provide adequate sputum to improve sputum collection for lung cancer diagnosis. PMID:26309391

  6. Analysis of Lung Flute-collected Sputum for Lung Cancer Diagnosis.

    PubMed

    Su, Jian; Anjuman, Nigar; Guarnera, Maria A; Zhang, Howard; Stass, Sanford A; Jiang, Feng

    2015-01-01

    Molecular analysis of sputum can help diagnose lung cancer. We have demonstrated that Lung Flute can be used to collect sputum from individuals who cannot spontaneously expectorate sputum. The objective of this study is to further evaluate the performance of the Lung Flute by comparing the characteristics of parallel samples collected with and without the Lung Flute and the usefulness for diagnosis of lung cancer. Fifty-six early-stage lung cancer patients (40 current smokers and 16 former smokers) and 73 cancer-free individuals (52 current smokers and 21 former smokers) were instructed to spontaneously cough and use Lung Flute for sputum sampling. Sputum cytology and polymerase chain reaction analysis of three miRNAs (miRs-21, 31, and 210) were performed in the specimens. All 92 current smokers and 11 (28.7%) of 37 former smokers spontaneously expectorated sputum and also produced sputum when using the Lung Flute. Twenty-seven former smokers (70.3%) who could not spontaneously expectorate sputum, however, were able to produce sputum when using the Lung Flute. The specimens were of low respiratory origin without contamination from other sources, eg, saliva. There was no difference of sputum volume and cell populations, diagnostic efficiency of cytology, and analysis of the miRNAs in the specimens collected by the two approaches. Analysis of the sputum miRNAs produced 83.93% sensitivity and 87.67% specificity for identifying lung cancer. Therefore, sputum collected by the Lung Flute has comparable features as spontaneously expectorated sputum. Using the Lung Flute enables former smokers who cannot spontaneously expectorate to provide adequate sputum to improve sputum collection for lung cancer diagnosis.

  7. Dryden B-52 Launch Aircraft on Edwards AFB Runway

    NASA Technical Reports Server (NTRS)

    1996-01-01

    NASA's venerable workhorse, the B-52 mothership, rolls out on the Edwards AFB runway after a test flight in 1996. Over the course of more than 40 years, the B-52 launched numerous experimental aircraft, ranging from the X-15 to the X-38, and was also used as a flying testbed for a variety of other research projects. NASA B-52, Tail Number 008, is an air launch carrier aircraft, 'mothership,' as well as a research aircraft platform that has been used on a variety of research projects. The aircraft, a 'B' model built in 1952 and first flown on June 11, 1955, is the oldest B-52 in flying status and has been used on some of the most significant research projects in aerospace history. Some of the significant projects supported by B-52 008 include the X-15, the lifting bodies, HiMAT (highly maneuverable aircraft technology), Pegasus, validation of parachute systems developed for the space shuttle program (solid-rocket-booster recovery system and the orbiter drag chute system), and the X-38. The B-52 served as the launch vehicle on 106 X-15 flights and flew a total of 159 captive-carry and launch missions in support of that program from June 1959 to October 1968. Information gained from the highly successful X-15 program contributed to the Mercury, Gemini, and Apollo human spaceflight programs as well as space shuttle development. Between 1966 and 1975, the B-52 served as the launch aircraft for 127 of the 144 wingless lifting body flights. In the 1970s and 1980s, the B-52 was the launch aircraft for several aircraft at what is now the Dryden Flight Research Center, Edwards, California, to study spin-stall, high-angle-of attack, and maneuvering characteristics. These included the 3/8-scale F-15/spin research vehicle (SRV), the HiMAT (Highly Maneuverable Aircraft Technology) research vehicle, and the DAST (drones for aerodynamic and structural testing). The aircraft supported the development of parachute recovery systems used to recover the space shuttle solid rocket

  8. Comparison of Integrated Gasifier-Combined Cycle and AFB-steam turbine systems for industrial cogeneration

    NASA Technical Reports Server (NTRS)

    Nainiger, J. J.; Abbott, J. M.; Burns, R. K.

    1981-01-01

    In the cogeneration technology alternatives study (CTAS) a number of advanced coal fired systems were examined and systems using a integrated coal gasifier IGCC or a fluid bed combustor AFB were found to yield attractive cogeneration results in industrial cogeneration applications. A range of site requirements and cogeneration sizing strategies using ground rules based on CTAS were used in comparing an IGCC and an AFB. The effect of time variations in site requirements and the sensitivity to fuel and electricity price assumptions are examined. The economic alternatives of industrial or utility ownership are also considered. The results indicate that the IGCC system has potentially higher fuel and emission savings and could be an attractive option for utility ownership. The AFB steam turbine system has a potentially higher return on investment and could be attractive assuming industrial ownership.

  9. Comparison of bronchial brushing and sputum in detection of pediatric pulmonary tuberculosis.

    PubMed

    Chen, Qiao-Pei; Ren, Shi-Feng; Wang, Xin-Feng; Wang, Mao-Shui

    2016-01-27

    The retrospective study aimed to evaluate the diagnostic value of bronchial brushing and sputum using acid fast bacilli smear, mycobacterial culture and real-time PCR in detection of pediatric pulmonary tuberculosis, sensitivity and specificity of bronchial brushing and sputum examined by the three methods were calculated and compared to each other. Data showed there were no significant difference in sensitivity between bronchial brushing and matched sputum using each method. But the specificity of real-time PCR on bronchial brushing was lower than on sputum. Compared with bronchial brushing, sputum was better specimen in detection of pediatric pulmonary tuberculosis.

  10. Sputum Induction in Children Is Feasible and Useful in a Bustling General Hospital Practice

    PubMed Central

    Bart, Ingeborg Y.; Mourits, Mariëlle; van Gent, René; van Leuken, Maarten H.; Hilbink, Mirrian; Warris, Adilia; Wever, Peter C.; de Vries, Esther

    2016-01-01

    We prospectively studied the feasibility and effectiveness of sputum induction in obtaining good quality sputum and its subsequent bacterial yield in children with clinically suspected acute lower-respiratory-tract infection (aLRTI). Good quality sputum was collected in 89/98 (91%) patients. Sputum cultures revealed ≥1 bacterial pathogens in 22 cases (25%). Adverse events were infrequent and mild (6%). Sputum induction is feasible in young children and leads to an increased number of etiological diagnoses of aLRTI. PMID:27336008

  11. Sputum RNA signature in allergic asthmatics following allergen bronchoprovocation test

    PubMed Central

    Zuiker, Rob G.J.A.; Tribouley, Catherine; Diamant, Zuzana; Boot, J. Diderik; Cohen, Adam F.; Van Dyck, K.; De Lepeleire, I.; Rivas, Veronica M.; Malkov, Vladislav A.; Burggraaf, Jacobus; Ruddy, Marcella K.

    2016-01-01

    Background Inhaled allergen challenge is a validated disease model of allergic asthma offering useful pharmacodynamic assessment of pharmacotherapeutic effects in a limited number of subjects. Objectives To evaluate whether an RNA signature can be identified from induced sputum following an inhaled allergen challenge, whether a RNA signature could be modulated by limited doses of inhaled fluticasone, and whether these gene expression profiles would correlate with the clinical endpoints measured in this study. Methods Thirteen non-smoking, allergic subjects with mild-to-moderate asthma participated in a randomised, placebo-controlled, 2-period cross-over study following a single-blind placebo run-in period. Each period consisted of three consecutive days, separated by a wash-out period of at least 3 weeks. Subjects randomly received inhaled fluticasone ((FP) MDI; 500 mcg BID×5 doses in total) or placebo. On day 2, house dust mite extract was inhaled and airway response was measured by FEV1 at predefined time points until 7 h post-allergen. Sputum was induced by NaCl 4.5%, processed and analysed at 24 h pre-allergen and 7 and 24 h post-allergen. RNA was isolated from eligible sputum cell pellets (<80% squamous of 500 cells), amplified according to NuGEN technology, and profiled on Affymetrix arrays. Gene expression changes from baseline and fluticasone treatment effects were evaluated using a mixed effects ANCOVA model at 7 and at 24 h post-allergen challenge. Results Inhaled allergen-induced statistically significant gene expression changes in sputum, which were effectively blunted by fluticasone (adjusted p<0.025). Forty-seven RNA signatures were selected from these responses for correlation analyses and further validation. This included Th2 mRNA levels for cytokines, chemokines, high-affinity IgE receptor FCER1A, histamine receptor HRH4, and enzymes and receptors in the arachidonic pathway. Individual messengers from the 47 RNA signatures correlated significantly

  12. In vitro study of AFB1 and AFM1 effects on human lymphoblastoid Jurkat T-cell model.

    PubMed

    Luongo, D; Russo, R; Balestrieri, A; Marzocco, S; Bergamo, P; Severino, L

    2014-10-01

    Aflatoxin B(1) (AFB(1)) is a mycotoxin produced by Aspergillus spp. that can occur as a natural contaminant in foods and feeds of vegetable origin. Post-ingestion, AFB(1) can be metabolized in the liver of mammals into hydroxylated aflatoxin M(1) (AFM(1)) that is excreted with milk. Although several studies have been carried out to evaluate effects of AFB(1) on the immune system, studies regarding AFM(1) are moreover lacking. The aim of the current study was to investigate effects of AFB(1) and AFM(1) on immune function using a lymphoblastoid Jurkat T-cell line as an experimental model. Both AFB(1) and AFM(1) produced significant decreases in Jurkat cell proliferation, whereas only minor effects were noted on interleukin (IL)-2 and interferon (IFN)-γ cytokines mRNA expression in stimulated cells that had been pre-incubated with AFB(1) and AFM(1). Particularly, AFB(1), but not AFM(1), at the highest concentration (50 µM) induced a marked increase in IL-8 mRNA expression. The results of the current study suggested the existence of a concentration threshold for AFB(1) and AFM(1) needed to exert biological activity on cell viability and innate immunity.

  13. Drone Transport of Microbes in Blood and Sputum Laboratory Specimens.

    PubMed

    Amukele, Timothy K; Street, Jeff; Carroll, Karen; Miller, Heather; Zhang, Sean X

    2016-10-01

    Unmanned aerial vehicles (UAVs) could potentially be used to transport microbiological specimens. To examine the impact of UAVs on microbiological specimens, blood and sputum culture specimens were seeded with usual pathogens and flown in a UAV for 30 ± 2 min. Times to recovery, colony counts, morphologies, and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based identifications of the flown and stationary specimens were similar for all microbes studied.

  14. 76 FR 38580 - Proposed Amendment of Class D Airspace; Eglin AFB, FL

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-01

    ... ``significant rule'' under DOT Regulatory Policies and Procedures (44 FR 11034; February 26, 1979); and (3) does..., 40120; E.O. 10854, 24 FR 9565, 3 CFR, 1959-1963 Comp., p. 389. Sec. 71.1 2. The incorporation by... Federal Aviation Administration 14 CFR Part 71 Proposed Amendment of Class D Airspace; Eglin AFB,...

  15. Studies on the biological functions of CPS1 in AFB1 induced hepatocarcinogenesis.

    PubMed

    Yang, Chi; Fu, Rao; Zhuang, Zhenhong; Wang, Shihua

    2016-10-10

    Carbamyl phosphate synthetase 1 (CPS1) was down-regulated in hepatocellular carcinoma (HCC), as treated by aflatoxin B1 (AFB1), a potent hepatocarcinogenesis mycotoxin. In this study, we firstly confirmed that AFB1 down-regulated the expression of CPS1 in a dose-dependent manner. At the meantime, both siRNA knock down of CPS1 and AFB1 treatment inhibited cell proliferation, and induced cell apoptosis. To further analysis the function of CPS1, the interacting proteins of CPS1 were searched by Co-IP, and three interacting proteins including type II cytoskeletal 1 (KRT1), albumin (ALB), and ubiquitin C (UBC) were found. Both KRT1 and ALB were new interacting proteins for CPS1. Our further study showed that CPS1 was regulating interacted and colocalized with KRT1 and ALB, and the intensity correlation was changed by AFB1. KRT1, ALB and CPS1 were all reported to play an important role in differentiation and tissue specialization. These results may offer an increasing understand that CPS1 might have a function in differentiation. PMID:27425868

  16. Response of a WB-47E Airplane to Runway Roughness at Eielson AFB, Alaska, September 1964

    NASA Technical Reports Server (NTRS)

    Morris, Garland J.; Hall, Albert W.

    1965-01-01

    An investigation has been conducted to measure the response of a WB-47E airplane to the roughness of the runway at Eielson AFB, Alaska. The acceleration level in the pilot's compartment and the pitching oscillation of the airplane were found to be sufficiently high to possibly cause pilot discomfort and have an adverse effect on the precision of take-off.

  17. 78 FR 33016 - Proposed Amendment of Class D Airspace; Grand Forks AFB, ND

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-03

    ... Regulatory Policies and Procedures (44 FR 11034; February 26, 1979); and (3) does not warrant preparation of... read as follows: Authority: 49 U.S.C. 106(g); 40103, 40113, 40120; E.O. 10854, 24 FR 9565, 3 CFR, 1959... Federal Aviation Administration 14 CFR Part 71 Proposed Amendment of Class D Airspace; Grand Forks AFB,...

  18. 33 CFR 334.748 - Wynnhaven Beach, Fla., at Eglin AFB; restricted area.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Beach, Fla., at Eglin AFB; restricted area. (a) The area. The restricted area shall encompass all navigable waters of the United States, as defined at 33 CFR part 329, within the area bounded by a line... 33 Navigation and Navigable Waters 3 2014-07-01 2014-07-01 false Wynnhaven Beach, Fla., at...

  19. 33 CFR 334.748 - Wynnhaven Beach, Fla., at Eglin AFB; restricted area.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Beach, Fla., at Eglin AFB; restricted area. (a) The area. The restricted area shall encompass all navigable waters of the United States, as defined at 33 CFR part 329, within the area bounded by a line... 33 Navigation and Navigable Waters 3 2013-07-01 2013-07-01 false Wynnhaven Beach, Fla., at...

  20. 33 CFR 334.748 - Wynnhaven Beach, Fla., at Eglin AFB; restricted area.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Beach, Fla., at Eglin AFB; restricted area. (a) The area. The restricted area shall encompass all navigable waters of the United States, as defined at 33 CFR part 329, within the area bounded by a line... 33 Navigation and Navigable Waters 3 2012-07-01 2012-07-01 false Wynnhaven Beach, Fla., at...

  1. Solvent-emissions reduction study at Newark AFB, Ohio. Final report, Aug 88-Apr 89

    SciTech Connect

    Ayer, J.; Wolbach, C.D.

    1990-05-01

    The objective of this effort was to collect baseline Freon emissions data, and subsequently recommend potential emission control alternatives to minimize Freon emissions that result from routine maintenance and repair operations conducted at Newark AFB, Ohio. Newark AFB, Ohio, uses a number of solvents to clean and maintain electronic guidance devices. The solvent most often used in this application is 1,1,2-trichloro-1,2,2-trifluoroethane, a solvent commonly known by the DuPont Company trademark Freon 113TM (hereafter referred to as Freon). Newark AFB purchases large quantities of Freon (nearly 600,000 pounds annually), and in previous years, lost nearly all of it (555,000 pounds) as unrecovered Freon vapor. Freon is one of a general class of chemicals known as chlorofluorocarbons (CFCs). Scientific evidence strongly suggests that CFC emissions are responsible for the depletion of the protective ozone layer surrounding the earth's atmosphere. For this reason, the U.S. air Force must reduce and eventually eliminate CFC emissions from Air Force facilities. Several steps have been taken to reduce the quantity of Freon emitted from the more than 100 emission point sources at Newark AFB. For example, the recovery of solvent vapors emitted from more than half of the point sources at the facility is achieved with limited success by the use of two carbon adsorption (CA) systems.

  2. Astronaut Jack R. Lousma in Water Survival Training near Homestead AFB

    NASA Technical Reports Server (NTRS)

    1978-01-01

    Three views documenting Astronaut Jack R. Lousma in Water Survival Training at the Air Force School at Turkey Point, near Homestead AFB. Views include long range view of Lousma parasailing, with tether attached to unseen craft (34035); view of Lousma in water with floatation devices attached to his arms (34036); close-up view of Lousma during parasailing training (34037).

  3. Sustainability of TCE Removal in the Mulch Biowalls at Altus AFB

    EPA Science Inventory

    A permeable mulch biowall was installed in June 2002 at Landfill 3 (LF-03), Operable Unit 1 (OU-1), Altus AFB, Oklahoma. The demonstration was conducted by Parsons for the AFCEE Technology Transfer Outreach Office. The biowall is approximately 455 feet long, by 24 feet deep, by...

  4. 77 FR 4457 - Amendment of Class D Airspace; Altus AFB, OK

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-30

    ... Federal Regulations is sold by the Superintendent of Documents. #0;Prices of new books are listed in the.... SUPPLEMENTARY INFORMATION: History On October 28, 2011, the FAA published in the Federal Register a notice of proposed rulemaking to amend Class D airspace for Altus AFB, OK (76 FR 66866) Docket No....

  5. STS-29 Discovery, OV-103, lands on Edwards AFB concrete runway 22

    NASA Technical Reports Server (NTRS)

    1989-01-01

    STS-29 Discovery, Orbiter Vehicle (OV) 103, main landing gear (MLG) touches down at a speed of approximately 205 knots (235 miles per hour) on concrete runway 22 at Edwards Air Force Base (AFB), California. Nose landing gear (NLG) is deployed and rides above runway surface prior touchdown. Mojave desert scrub brush appears in the foreground with mountain range appearing in the background.

  6. 76 FR 66866 - Proposed Amendment of Class D Airspace; Altus AFB, OK

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-28

    ...'' under DOT Regulatory Policies and Procedures (44 FR 11034; February 26, 1979); and (3) does not warrant...: Authority: 49 U.S.C. 106(g); 40103, 40113, 40120; E.O. 10854, 24 FR 9565, 3 CFR, 1959-1963 Comp., p. 389... Federal Aviation Administration 14 CFR Part 71 Proposed Amendment of Class D Airspace; Altus AFB,...

  7. Sputum colour and bacteria in chronic bronchitis exacerbations: a pooled analysis.

    PubMed

    Miravitlles, Marc; Kruesmann, Frank; Haverstock, Daniel; Perroncel, Renee; Choudhri, Shurjeel H; Arvis, Pierre

    2012-06-01

    We examined the correlation between sputum colour and the presence of potentially pathogenic bacteria in acute exacerbations of chronic bronchitis (AECBs). Data were pooled from six multicentre studies comparing moxifloxacin with other antimicrobials in patients with an AECB. Sputum was collected before antimicrobial therapy, and bacteria were identified by culture and Gram staining. Association between sputum colour and bacteria was determined using logistic regression. Of 4,089 sputum samples, a colour was reported in 4,003; 1,898 (46.4%) were culture-positive. Green or yellow sputum samples were most likely to yield bacteria (58.9% and 45.5% of samples, respectively), compared with 18% of clear and 39% of rust-coloured samples positive for potentially pathogenic microorganisms. Factors predicting a positive culture were sputum colour (the strongest predictor), sputum purulence, increased dyspnoea, male sex and absence of fever. Green or yellow versus white sputum colour was associated with a sensitivity of 94.7% and a specificity of 15% for the presence of bacteria. Sputum colour, particularly green and yellow, was a stronger predictor of potentially pathogenic bacteria than sputum purulence and increased dyspnoea in AECB patients. However, it does not necessarily predict the need for antibiotic treatment in all patients with AECB.

  8. Effect of expectoration on inflammation in induced sputum in alpha-1-antitrypsin deficiency.

    PubMed

    Gompertz, Simon; Hill, Adam T; Bayley, Darren L; Stockley, Robert A

    2006-06-01

    It is unclear how chronic expectoration influences airway inflammation in patients with chronic lung disease. The aim of this study was to investigate factors influencing inflammation in induced sputum samples, including, in particular, chronic sputum production. Myeloperoxidase, interleukin-8, leukotriene B4 (LTB4), neutrophil elastase, secretory leukoprotease inhibitor (SLPI) and protein leakage were compared in induced sputum samples from 48 patients (36 with chronic expectoration) with COPD (with and without alpha-1-antitrypsin deficiency; AATD), 9 individuals with AATD but without lung disease and 14 healthy controls. There were no differences in inflammation in induced sputum samples from healthy control subjects and from AATD deficient patients with normal lung function but without chronic expectoration (P>0.05). Inflammation in induced sputum from AATD patients with airflow obstruction and chronic sputum expectoration was significantly greater than for similar patients who did not expectorate: Interleukin-8 (P<0.01), elastase activity (P=0.01), and protein leakage (P<0.01). The presence of spontaneous sputum expectoration in AATD patients with airflow obstruction was associated with increased neutrophilic airway inflammation in induced sputum samples. The presence of chronic expectoration in some patients will clearly complicate interpretation of studies employing sputum induction where this feature has not been identified.

  9. Protein networks in induced sputum from smokers and COPD patients

    PubMed Central

    Baraniuk, James N; Casado, Begona; Pannell, Lewis K; McGarvey, Peter B; Boschetto, Piera; Luisetti, Maurizio; Iadarola, Paolo

    2015-01-01

    Rationale Subtypes of cigarette smoke-induced disease affect different lung structures and may have distinct pathophysiological mechanisms. Objective To determine if proteomic classification of the cellular and vascular origins of sputum proteins can characterize these mechanisms and phenotypes. Subjects and methods Individual sputum specimens from lifelong nonsmokers (n=7) and smokers with normal lung function (n=13), mucous hypersecretion with normal lung function (n=11), obstructed airflow without emphysema (n=15), and obstruction plus emphysema (n=10) were assessed with mass spectrometry. Data reduction, logarithmic transformation of spectral counts, and Cytoscape network-interaction analysis were performed. The original 203 proteins were reduced to the most informative 50. Sources were secretory dimeric IgA, submucosal gland serous and mucous cells, goblet and other epithelial cells, and vascular permeability. Results Epithelial proteins discriminated nonsmokers from smokers. Mucin 5AC was elevated in healthy smokers and chronic bronchitis, suggesting a continuum with the severity of hypersecretion determined by mechanisms of goblet-cell hyperplasia. Obstructed airflow was correlated with glandular proteins and lower levels of Ig joining chain compared to other groups. Emphysema subjects’ sputum was unique, with high plasma proteins and components of neutrophil extracellular traps, such as histones and defensins. In contrast, defensins were correlated with epithelial proteins in all other groups. Protein-network interactions were unique to each group. Conclusion The proteomes were interpreted as complex “biosignatures” that suggest distinct pathophysiological mechanisms for mucin 5AC hypersecretion, airflow obstruction, and inflammatory emphysema phenotypes. Proteomic phenotyping may improve genotyping studies by selecting more homogeneous study groups. Each phenotype may require its own mechanistically based diagnostic, risk-assessment, drug- and other

  10. The Sputum Microbiome in Chronic Obstructive Pulmonary Disease Exacerbations.

    PubMed

    Huang, Yvonne J; Boushey, Homer A

    2015-11-01

    Acute exacerbations of chronic obstructive pulmonary disease (COPD) are thought to be associated with--and perhaps to mediate--accelerated loss of lung function in COPD. Although the application of culture-independent methods for detection of bacteria have shown COPD to be associated with marked differences in the burden, diversity, and composition of the bronchial bacterial microbiome, few studies have examined the changes associated with community-acquired exacerbations of the disease. In a longitudinal cohort study of COPD, the availability of sputum samples from subjects obtained at the onset of an exacerbation and during periods of clinical stability before and after the event enabled us to recently address this gap in knowledge, using culture-independent, 16S rRNA-based analysis methods combined with in silico inference of metagenomic functions. We observed sputum bacterial composition to be generally stable over the preexacerbation period of clinical stability, but to change at the time of exacerbation, with specific enrichment in not only typical COPD-associated bacterial species (e.g., Haemophilus influenzae) but also other phylogenetically related species with pathogenic potential. Concurrently, we observed depleted abundance of other bacteria whose predicted metagenomes suggest functional capacities to produce a variety of antiinflammatory compounds. Most strikingly, we found that resolution of these exacerbation-related changes in sputum microbiota composition differed significantly, depending on the exacerbation treatments prescribed. Treatment with corticosteroids resulted in microbiome enrichment for a number of bacterial communities, mostly members of the Proteobacteria phylum, whereas prolonged suppression of microbiota was seen in those treated with antibiotics alone. Taken together, our findings suggest that exacerbations of COPD are associated with heterogeneous changes in the bronchial microbiome, with increases in the abundance of species

  11. The Sputum Microbiome in Chronic Obstructive Pulmonary Disease Exacerbations.

    PubMed

    Huang, Yvonne J; Boushey, Homer A

    2015-11-01

    Acute exacerbations of chronic obstructive pulmonary disease (COPD) are thought to be associated with--and perhaps to mediate--accelerated loss of lung function in COPD. Although the application of culture-independent methods for detection of bacteria have shown COPD to be associated with marked differences in the burden, diversity, and composition of the bronchial bacterial microbiome, few studies have examined the changes associated with community-acquired exacerbations of the disease. In a longitudinal cohort study of COPD, the availability of sputum samples from subjects obtained at the onset of an exacerbation and during periods of clinical stability before and after the event enabled us to recently address this gap in knowledge, using culture-independent, 16S rRNA-based analysis methods combined with in silico inference of metagenomic functions. We observed sputum bacterial composition to be generally stable over the preexacerbation period of clinical stability, but to change at the time of exacerbation, with specific enrichment in not only typical COPD-associated bacterial species (e.g., Haemophilus influenzae) but also other phylogenetically related species with pathogenic potential. Concurrently, we observed depleted abundance of other bacteria whose predicted metagenomes suggest functional capacities to produce a variety of antiinflammatory compounds. Most strikingly, we found that resolution of these exacerbation-related changes in sputum microbiota composition differed significantly, depending on the exacerbation treatments prescribed. Treatment with corticosteroids resulted in microbiome enrichment for a number of bacterial communities, mostly members of the Proteobacteria phylum, whereas prolonged suppression of microbiota was seen in those treated with antibiotics alone. Taken together, our findings suggest that exacerbations of COPD are associated with heterogeneous changes in the bronchial microbiome, with increases in the abundance of species

  12. ASCAN Precourt floats on life raft during Elgin AFB water survival training

    NASA Technical Reports Server (NTRS)

    1990-01-01

    1990 Group 13 Astronaut Candidate (ASCAN) Charles J. Precourt, wearing helmet and flight suit, floats in pool using an underarm flotation device and a single person life raft at Elgin Air Force Base (AFB) in Pensacola, Florida, during water survival exercises. The training familiarized the candidates with survival techniques necessary in the event of a water landing. ASCANs participated in the exercises from 08-14-90 through 08-17-90.

  13. Comparison of two methods of collecting induced sputum in asthmatic subjects.

    PubMed

    Gershman, N H; Wong, H H; Liu, J T; Mahlmeister, M J; Fahy, J V

    1996-12-01

    The method that we have previously reported for sputum induction involves collecting the entire expectorate produced over a 20 min inhalation of 3% saline aerosol. This method presents the potential disadvantage of a considerable and variable salivary contribution to the induced sputum sample. In this study, we examined whether separate collection of saliva and sputum represents a better method for collecting induced sputum during sputum induction. In 11 stable asthmatics, we compared the volume, total and differential cell counts, and eosinophil cationic protein (ECP) levels in four induced sputum samples, two performed using our previous method (Method A) and two using another method (Method B) in which subjects spit saliva into one container before coughing sputum into another. We found that the volume of sputum obtained with Method B was lower than that obtained with Method A (6.16 +/- 0.61 vs 20.1 +/- 2.7 mL; p = 0.003), as was the percentage of squamous cells (34 +/- 4 vs 47 +/- 6; p = 0.023). In addition, the ECP levels in samples collected by Method B were higher (261 +/- 42 vs 145 +/- 26 ng.mL-1; p = 0.01). The differential counts of nonsquamous cells were similar except for the percentage of neutrophils, which was lower in Method B (37 +/- 4 vs 50 +/- 5%; p = 0.019). The repeatability of measurements of eosinophil percentages and of ECP levels was similar for the two methods. We conclude that separate collection of saliva and sputum yields induced sputum samples with reduced amounts of saliva and is, therefore, a better method for collecting induced sputum.

  14. Bacterial infection in exacerbated COPD with changes in sputum characteristics.

    PubMed Central

    Monsó, E.; Garcia-Aymerich, J.; Soler, N.; Farrero, E.; Felez, M. A.; Antó, J. M.; Torres, A.

    2003-01-01

    We examined the risk factors for bacterial exacerbation, defined as the presence of pathogenic bacteria in sputum, in 90 chronic obstructive pulmonary disease (COPD) patients with an exacerbation and changes in sputum characteristics. Smoking, alcohol, lung function, body mass index, medical visits and treatments were the independent variables assessed using multivariable logistic regression modelling (OR, 95% CI). A bacterial exacerbation was diagnosed in 39 (43.3%) of 90 patients. Bacterial exacerbations were more prevalent among current smokers (OR 3.77, 95% CI 1.17-12.12), in patients with poor compliance with inhalation therapy (OR 3.25, 95% CI 1.18-8.93) and with severe lung function impairment (FEV1 OR 0.96, 95% CI 0.93-1.00). Prior use of antibiotics was a risk factor for Pseudomonas aeruginosa infection (OR 6.06, 95% CI 1.29-28.44) and influenza vaccination appeared to have a protective effect against this infection (OR 0.15, 95% CI 0.03-0.67). We conclude that severe impairment of lung function, smoking and poor compliance with therapy are risk factors for bacterial infection in COPD, and P. aeruginosa should be suspected in patients who have been treated with antibiotics and in those not vaccinated against influenza. PMID:12948381

  15. Environmental arsenic exposure and microbiota in induced sputum.

    PubMed

    White, Allison G; Watts, George S; Lu, Zhenqiang; Meza-Montenegro, Maria M; Lutz, Eric A; Harber, Philip; Burgess, Jefferey L

    2014-02-21

    Arsenic exposure from drinking water is associated with adverse respiratory outcomes, but it is unknown whether arsenic affects pulmonary microbiota. This exploratory study assessed the effect of exposure to arsenic in drinking water on bacterial diversity in the respiratory tract of non-smokers. Induced sputum was collected from 10 subjects with moderate mean household water arsenic concentration (21.1 ± 6.4 ppb) and 10 subjects with low household water arsenic (2.4 ± 0.8 ppb). To assess microbiota in sputum, the V6 hypervariable region amplicons of bacterial 16s rRNA genes were sequenced using the Ion Torrent Personal Genome Machine. Microbial community differences between arsenic exposure groups were evaluated using QIIME and Metastats. A total of 3,920,441 sequence reads, ranging from 37,935 to 508,787 per sample for 316 chips after QIIME quality filtering, were taxonomically classified into 142 individual genera and five phyla. Firmicutes (22%), Proteobacteria (17%) and Bacteriodetes (12%) were the main phyla in all samples, with Neisseriaceae (15%), Prevotellaceae (12%) and Veillonellacea (7%) being most common at the genus level. Some genera, including Gemella, Lactobacillales, Streptococcus, Neisseria and Pasteurellaceae were elevated in the moderate arsenic exposure group, while Rothia, Prevotella, Prevotellaceae Fusobacterium and Neisseriaceae were decreased, although none of these differences was statistically significant. Future studies with more participants and a greater range of arsenic exposure are needed to further elucidate the effects of drinking water arsenic consumption on respiratory microbiota.

  16. Environmental Arsenic Exposure and Microbiota in Induced Sputum

    PubMed Central

    White, Allison G.; Watts, George S.; Lu, Zhenqiang; Meza-Montenegro, Maria M.; Lutz, Eric A.; Harber, Philip; Burgess, Jefferey L.

    2014-01-01

    Arsenic exposure from drinking water is associated with adverse respiratory outcomes, but it is unknown whether arsenic affects pulmonary microbiota. This exploratory study assessed the effect of exposure to arsenic in drinking water on bacterial diversity in the respiratory tract of non-smokers. Induced sputum was collected from 10 subjects with moderate mean household water arsenic concentration (21.1 ± 6.4 ppb) and 10 subjects with low household water arsenic (2.4 ± 0.8 ppb). To assess microbiota in sputum, the V6 hypervariable region amplicons of bacterial 16s rRNA genes were sequenced using the Ion Torrent Personal Genome Machine. Microbial community differences between arsenic exposure groups were evaluated using QIIME and Metastats. A total of 3,920,441 sequence reads, ranging from 37,935 to 508,787 per sample for 316 chips after QIIME quality filtering, were taxonomically classified into 142 individual genera and five phyla. Firmicutes (22%), Proteobacteria (17%) and Bacteriodetes (12%) were the main phyla in all samples, with Neisseriaceae (15%), Prevotellaceae (12%) and Veillonellacea (7%) being most common at the genus level. Some genera, including Gemella, Lactobacillales, Streptococcus, Neisseria and Pasteurellaceae were elevated in the moderate arsenic exposure group, while Rothia, Prevotella, Prevotellaceae Fusobacterium and Neisseriaceae were decreased, although none of these differences was statistically significant. Future studies with more participants and a greater range of arsenic exposure are needed to further elucidate the effects of drinking water arsenic consumption on respiratory microbiota. PMID:24566055

  17. Pseudomonas siderophores in the sputum of patients with cystic fibrosis.

    PubMed

    Martin, Lois W; Reid, David W; Sharples, Katrina J; Lamont, Iain L

    2011-12-01

    The lungs of patients with cystic fibrosis become chronically infected with the bacterium Pseudomonas aeruginosa, which heralds progressive lung damage and a decline in health. Iron is a crucial micronutrient for bacteria and its acquisition is a key factor in infection. P. aeruginosa can acquire this element by secreting pyoverdine and pyochelin, iron-chelating compounds (siderophores) that scavenge iron and deliver it to the bacteria. Siderophore-mediated iron uptake is generally considered a key factor in the ability of P. aeruginosa to cause infection. We have investigated the amounts of pyoverdine in 148 sputum samples from 36 cystic fibrosis patients (30 infected with P. aeruginosa and 6 as negative controls). Pyoverdine was present in 93 samples in concentrations between 0.30 and 51 μM (median 4.6 μM) and there was a strong association between the amount of pyoverdine and the number of P. aeruginosa present. However, pyoverdine was not present, or below the limits of detection (~0.3 μM), in 21 sputum samples that contained P. aeruginosa. Pyochelin was also absent, or below the limits of detection (~1 μM), in samples from P. aeruginosa-infected patients with little or no detectable pyoverdine. Our data show that pyoverdine is an important iron-scavenging molecule for P. aeruginosa in many cystic fibrosis patients, but other P. aeruginosa iron-uptake systems must be active in some patients to satisfy the bacterial need for iron. PMID:21643731

  18. Bioventing pilot test results at the low point drain area, Offutt AFB, Nebraska.

    PubMed

    Werner, F T; Walters, J E; Keefer, G B

    1997-11-21

    The purpose of this paper was to describe the application of bioventing technology at the LPD site at Offutt AFB, Nebraska and present the results of the 15-month pilot test. The preliminary tests indicated sufficient hydrocarbon contamination was present with the necessary soil characteristics to warrant an extended bioventing pilot test. The six month in situ respiration test indicated that progress was being made in reducing the TVH concentrations and biological activity was still occurring. Laboratory analysis of the final soil samples confirmed the reduction in TRPH and BTEX concentrations indicating that the site is close to complete remediation. However, owing to reduced air flow at greater distances from the VW, more biodegradation is still needed near MPB. The reduced biodegradation at MPB could also be due to the high water tables resulting from heavy rains during the summer and fall of 1993. The local water table was above the VW and MP screens for several months. The operation of the blower will continue until the site is completely remediated. The single VW pilot test at the LPD site at Offutt AFB has proven the effectiveness of bioventing in reducing TRPH and BTEX contamination in the subsurface. The installation, operation and maintenance costs were minimal. The effectiveness of this application has resulted in three additional bioventing applications at Offutt AFB including the first, full-scale system located in the state of Nebraska.

  19. Flow cytometry of sputum: assessing inflammation and immune response elements in the bronchial airways**

    EPA Science Inventory

    Rationale: The evaluation of sputum leukocytes by flow cytometry is an opportunity to assess characteristics of cells residing in the central airways, yet it is hampered by certain inherent properties of sputum including mucus and large amounts of contaminating cells and debris. ...

  20. Sputum is a surrogate for bronchoalveolar lavage for monitoring Mycobacterium tuberculosis transcriptional profiles in TB patients.

    PubMed

    Garcia, Benjamin J; Loxton, Andre G; Dolganov, Gregory M; Van, Tran T; Davis, J Lucian; de Jong, Bouke C; Voskuil, Martin I; Leach, Sonia M; Schoolnik, Gary K; Walzl, Gerhard; Strong, Michael; Walter, Nicholas D

    2016-09-01

    Pathogen-targeted transcriptional profiling in human sputum may elucidate the physiologic state of Mycobacterium tuberculosis (M. tuberculosis) during infection and treatment. However, whether M. tuberculosis transcription in sputum recapitulates transcription in the lung is uncertain. We therefore compared M. tuberculosis transcription in human sputum and bronchoalveolar lavage (BAL) samples from 11 HIV-negative South African patients with pulmonary tuberculosis. We additionally compared these clinical samples with in vitro log phase aerobic growth and hypoxic non-replicating persistence (NRP-2). Of 2179 M. tuberculosis transcripts assayed in sputum and BAL via multiplex RT-PCR, 194 (8.9%) had a p-value <0.05, but none were significant after correction for multiple testing. Categorical enrichment analysis indicated that expression of the hypoxia-responsive DosR regulon was higher in BAL than in sputum. M. tuberculosis transcription in BAL and sputum was distinct from both aerobic growth and NRP-2, with a range of 396-1020 transcripts significantly differentially expressed after multiple testing correction. Collectively, our results indicate that M. tuberculosis transcription in sputum approximates M. tuberculosis transcription in the lung. Minor differences between M. tuberculosis transcription in BAL and sputum suggested lower oxygen concentrations or higher nitric oxide concentrations in BAL. M. tuberculosis-targeted transcriptional profiling of sputa may be a powerful tool for understanding M. tuberculosis pathogenesis and monitoring treatment responses in vivo.

  1. Comparison of expectorated sputum after manual chest physical therapy and high-frequency chest compression.

    PubMed

    Warwick, Warren J; Wielinski, Catherine L; Hansen, Leland G

    2004-01-01

    This study is a quantitative comparison of the sputum produced by 12 subjects with cystic fibrosis (CF) who received high-frequency chest compression (HFCC) and standard chest physical therapy (CPT) in randomized order. Six subjects routinely used manual CPT and six routinely used the HFCC. None had acute infections or hospitalization in the six weeks before the study. Two certified respiratory therapists alternated subjects and CPT vs HFCC order during the two weeks of the matched study. For all sessions, the expectorated sputum was collected in preweighed cups, which were reweighed immediately after collection and again after evaporation to dryness. The wet and dry weights of the sputum produced as a result of the two techniques were significantly different, with HFCC having greater weight. Regardless of the mode of therapy, the sputum produced by the subjects who regularly received HFCC had greater water content than did the sputum produced by those subjects who regularly received CPT. No significant difference was found between the two therapists regarding sputum expectorated by the subjects during CPT. These results show that sputum production by subjects with CF who receive CPT by certified respiratory therapists can be as great as the sputum produced by the same subjects who receive HFCC. The results also suggest that unknown factors attributed to the therapists may produce different levels of effort from time to time that may decrease the respiratory therapists' effectiveness, whereas the HFCC therapy may be more consistently effective because it is entirely machine based.

  2. Sputum is a surrogate for bronchoalveolar lavage for monitoring Mycobacterium tuberculosis transcriptional profiles in TB patients.

    PubMed

    Garcia, Benjamin J; Loxton, Andre G; Dolganov, Gregory M; Van, Tran T; Davis, J Lucian; de Jong, Bouke C; Voskuil, Martin I; Leach, Sonia M; Schoolnik, Gary K; Walzl, Gerhard; Strong, Michael; Walter, Nicholas D

    2016-09-01

    Pathogen-targeted transcriptional profiling in human sputum may elucidate the physiologic state of Mycobacterium tuberculosis (M. tuberculosis) during infection and treatment. However, whether M. tuberculosis transcription in sputum recapitulates transcription in the lung is uncertain. We therefore compared M. tuberculosis transcription in human sputum and bronchoalveolar lavage (BAL) samples from 11 HIV-negative South African patients with pulmonary tuberculosis. We additionally compared these clinical samples with in vitro log phase aerobic growth and hypoxic non-replicating persistence (NRP-2). Of 2179 M. tuberculosis transcripts assayed in sputum and BAL via multiplex RT-PCR, 194 (8.9%) had a p-value <0.05, but none were significant after correction for multiple testing. Categorical enrichment analysis indicated that expression of the hypoxia-responsive DosR regulon was higher in BAL than in sputum. M. tuberculosis transcription in BAL and sputum was distinct from both aerobic growth and NRP-2, with a range of 396-1020 transcripts significantly differentially expressed after multiple testing correction. Collectively, our results indicate that M. tuberculosis transcription in sputum approximates M. tuberculosis transcription in the lung. Minor differences between M. tuberculosis transcription in BAL and sputum suggested lower oxygen concentrations or higher nitric oxide concentrations in BAL. M. tuberculosis-targeted transcriptional profiling of sputa may be a powerful tool for understanding M. tuberculosis pathogenesis and monitoring treatment responses in vivo. PMID:27553415

  3. [Determining asthma treatment in children by monitoring fractional exhaled nitric oxide, sputum eosinophils and leukotriene B₄].

    PubMed

    Vizmanos-Lamotte, G; Cruz, M J; Gómez-Ollés, S; Muñoz, X; de Mir Messa, I; Moreno-Galdó, A

    2015-01-01

    Sputum eosinophils and exhaled fractional nitric oxide (FENO) are markers of airway inflammation in asthma. Cytokines, cysteinyl-leukotrienes and leukotriene B4 (LTB4) are responsible for this inflammation. The aim of this study is to determine the usefulness of these markers in monitoring asthma treatment in children. FENO, sputum eosinophils, and LTB4 in induced sputum were performed in 10 children (9-15 years old). These determinations were repeated four months later, after the beginning or an increase in the treatment. FENO values tended to decrease (P=.15), pulmonary function tended to improve (P=.10), and sputum eosinophils decreased (P=.003) compared to the first determination. There were no differences in LTB4 concentrations (P=.88). Sputum eosinophils seem to be more precise than FENO in the monitoring of inflammation in asthmatic children.

  4. Comparison of sputum and nasopharyngeal swabs for detection of respiratory viruses.

    PubMed

    Jeong, Ji Hun; Kim, Kyung Hee; Jeong, Sung Hwan; Park, Jeong Woong; Lee, Sang Min; Seo, Yiel Hea

    2014-12-01

    Diagnostic tests for respiratory viral infections use traditionally either nasopharyngeal washes or swabs. Sputum is representative of the lower respiratory tract but is used rarely for viral testing. The aim of this study was to compare the detection rates of respiratory viruses from nasopharyngeal swabs and sputum using a multiplex real-time reverse transcription-polymerase chain reaction (RT-PCR). Adults who were admitted or presented to the clinics of Gil Medical Center with acute respiratory symptoms were recruited from 1 November 2012 to 31 March 2013. Paired specimens of nasopharyngeal swabs and sputum were obtained from 154 subjects, and RNA was extracted and tested for 16 different respiratory viruses using the Anyplex II RV16 Detection kit (Seegene, Seoul, Korea). The positive rate was 53% (81/154) for nasopharyngeal swabs and 68% (105/154) for sputum (P < 0.001). One hundred thirty-four viruses were identified for 107 illnesses. Influenza A virus, RSV A, HRV, coronavirus OC43, and adenovirus were detected more frequently in sputum samples than in nasopharyngeal swabs (P < 0.001). Importantly, 12 of 44 (27%) influenza A infections and 11 of 27 (41%) RSV infections were positive in only sputum samples. The detection rates of respiratory viruses from sputum samples were significantly higher than those from nasopharyngeal swabs in adults using real-time multiplex RT-PCR. These findings suggest that sputum would benefit for the detection of respiratory viruses by nucleic acid amplification tests (NAATs) in patients who produce sputum. Further studies are needed to establish standardized RNA extraction methods from sputum samples.

  5. Identification of mycobacterium tuberculosis in sputum smear slide using automatic scanning microscope

    NASA Astrophysics Data System (ADS)

    Rulaningtyas, Riries; Suksmono, Andriyan B.; Mengko, Tati L. R.; Saptawati, Putri

    2015-04-01

    Sputum smear observation has an important role in tuberculosis (TB) disease diagnosis, because it needs accurate identification to avoid high errors diagnosis. In development countries, sputum smear slide observation is commonly done with conventional light microscope from Ziehl-Neelsen stained tissue and it doesn't need high cost to maintain the microscope. The clinicians do manual screening process for sputum smear slide which is time consuming and needs highly training to detect the presence of TB bacilli (mycobacterium tuberculosis) accurately, especially for negative slide and slide with less number of TB bacilli. For helping the clinicians, we propose automatic scanning microscope with automatic identification of TB bacilli. The designed system modified the field movement of light microscope with stepper motor which was controlled by microcontroller. Every sputum smear field was captured by camera. After that some image processing techniques were done for the sputum smear images. The color threshold was used for background subtraction with hue canal in HSV color space. Sobel edge detection algorithm was used for TB bacilli image segmentation. We used feature extraction based on shape for bacilli analyzing and then neural network classified TB bacilli or not. The results indicated identification of TB bacilli that we have done worked well and detected TB bacilli accurately in sputum smear slide with normal staining, but not worked well in over staining and less staining tissue slide. However, overall the designed system can help the clinicians in sputum smear observation becomes more easily.

  6. Profiling over 1500 lipids in induced lung sputum and the implications in studying lung diseases.

    PubMed

    t'Kindt, Ruben; Telenga, Eef D; Jorge, Lucie; Van Oosterhout, Antoon J M; Sandra, Pat; Ten Hacken, Nick H T; Sandra, Koen

    2015-01-01

    Induced lung sputum is a valuable matrix in the study of respiratory diseases. Although the methodology of sputum collection has evolved to a point where it is repeatable and responsive to inflammation, its use in molecular profiling studies is still limited. Here, an in-depth lipid profiling of induced lung sputum using high-resolution liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (LC-Q-TOF MS) is described. An enormous complexity in lipid composition could be revealed. Over 1500 intact lipids, originating from 6 major lipid classes, have been accurately identified in 120 μL of induced sputum. By number and measured intensity, glycerophospholipids represent the largest lipid class, followed by sphingolipids, glycerolipids, fatty acyls, sterol lipids, and prenol lipids. Several prenol lipids, originating from tobacco, could be detected in the lung sputum of smokers. To illustrate the utility of the methodology in studying respiratory diseases, a comparative lipid screening was performed on lung sputum extracts in order to study the effect of Chronic Obstructive Pulmonary Disease (COPD) on the lung barrier lipidome. Results show that sphingolipid expression in induced sputum significantly differs between smokers with and without COPD.

  7. Group 13, 1990 ASCAN Cockrell in ejection seat trainer at Vance AFB, Oklahoma

    NASA Technical Reports Server (NTRS)

    1990-01-01

    Group 13, 1990 Astronaut Candidate (ASCAN) Kenneth D. Cockrell, wearing flight suit and helmet, anticipates the activation of the ejection seat on which he is sitting. He is participating in one of many sessions conducted during a survival training course at Vance Air Force Base (AFB) in Enid, Oklahoma. This portion of the survival training course is designed to familiarize the trainee with the 'feel' of emergency ejection from a jet aircraft. Cockrell, along with 22 other ASCANs, began a year's training and evaluation in July. This session was conducted from 07-29-90 through 07-31-90.

  8. ASCAN Ochoa floats in pool during Elgin AFB water survival training

    NASA Technical Reports Server (NTRS)

    1990-01-01

    1990 Group 13 Astronaut Candidate (ASCAN) Ellen Ochoa, wearing helmet and flight suit, climbs into a single person life raft while floating in a pool at Elgin Air Force Base (AFB) in Pensacola, Florida, during water survival exercises. Ochoa's underarm flotation device holds her above the water as she pulls herself into the life raft. The training familiarized the candidates with survival techniques necessary in the event of a water landing. ASCANs participated in the exercises from 08-14-90 through 08-17-90.

  9. STS-29 Discovery, OV-103, lands on Edwards AFB concrete runway 22

    NASA Technical Reports Server (NTRS)

    1989-01-01

    STS-29 Discovery, Orbiter Vehicle (OV) 103, main landing gear (MLG) touches down at a speed of approximately 205 knots (235 miles per hour) on concrete runway 22 at Edwards Air Force Base (AFB), California. Nose landing gear (NLG) is deployed and rides above runway surface prior touchdown. Rear view captures OV-103 as it glides past photographer to wheel stop showing the tail section (speedbrake/rudder) and three space shuttle main engines (SSMEs). Mojave desert scrub brush appears in the foreground with aircraft hangar appearing in the background.

  10. Rhinitis Patients With Sputum Eosinophilia Show Decreased Lung Function in the Absence of Airway Hyperresponsiveness

    PubMed Central

    Yang, Min-Suk; Lee, Hyun-Seung; Kim, Min-Hye; Song, Woo-Jung; Kim, Tae-Wan; Kwon, Jae-Woo; Kim, Sae-Hoon; Park, Heung-Woo; Chang, Yoon-Seok; Min, Kyung-Up

    2013-01-01

    Purpose Sputum eosinophilia is observed frequently in patients with rhinitis. Sputum eosinophilia in patients with non-asthmatic allergic rhinitis has been suggested to be related to nonspecific airway hyperresponsiveness (AHR). However, the clinical significance of sputum eosinophilia in patients with non-asthmatic rhinitis without AHR has not been determined. We conducted a retrospective study examining the influence of sputum eosinophilia in patients with non-asthmatic rhinitis without AHR on pulmonary function and expression of fibrosis-related mediators. Methods Eighty-nine patients with moderate-to-severe perennial rhinitis without AHR were included. All underwent lung function tests (forced expiratory volume in 1 second [FEV1] and forced vital capacity [FVC]), skin tests to inhalant allergens, methacholine bronchial challenge tests, and hypertonic saline-induced sputum to determine eosinophil counts. Sputum mRNA levels for transforming growth factor-β (TGF-β), matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of metalloproteinase-1 (TIMP-1) were also examined. Patients were divided into two groups according to the presence of sputum eosinophilia (≥3%, eosinophilia-positive [EP] and <3%, eosinophilia-negative [EN] groups). Results FEV1 was significantly lower (P=0.04) and FEV1/FVC tended to be lower (P=0.1) in the EP group than in the EN group. In sputum analyses, the MMP-9 mRNA level (P=0.005) and the ratio of MMP-9 to TIMP-1 expression (P=0.01) were significantly higher in the EP group than in the EN group. There was no significant difference in TGF-β mRNA expression between the two groups. Conclusions Sputum eosinophilia in patients with moderate-to-severe perennial rhinitis without AHR influenced FEV1 and the expression pattern of fibrosis-related mediators. PMID:23814677

  11. Cystic fibrosis and bacterial colonization define the sputum N-glycosylation phenotype.

    PubMed

    Venkatakrishnan, Vignesh; Thaysen-Andersen, Morten; Chen, Sharon C A; Nevalainen, Helena; Packer, Nicolle H

    2015-01-01

    Although mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) is known to be altered relative to their unaffected counterparts, protein N-glycosylation of CF sputum remains structurally and functionally under-characterized. We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. N-Glycans were profiled using porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry following enzymatic release from sputum proteins. The composition, topology and linkage isomers of 68 N-glycans were characterized and relatively quantified. Recurring structural features in all sputum N-glycomes were terminal α2,6-sialylation, α1,6-core fucosylation, β1,4-bisecting GlcNAcylation and compositions indicating paucimannosylation. Despite covering different genotypes, age, gender and microbial flora, the sputum N-glycomes showed little interpatient and longitudinal variation within CF patients. Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. In contrast, the sputum from CF patients had only slightly increased abundance of paucimannose N-glycans relative to pathogen-infected/colonized non-CF individuals. Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. This study provides new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract. PMID:25190359

  12. Acid-fast bacilli in sputum: a case of Legionella micdadei pneumonia.

    PubMed Central

    Hilton, E; Freedman, R A; Cintron, F; Isenberg, H D; Singer, C

    1986-01-01

    Legionella micdadei has been implicated as a cause of nosocomial pneumonia. There are no reports of L. micdadei pneumonia diagnosed by acid-fast stain of expectorated sputum. We report a case of L. micdadei pneumonia in which expectorated sputum harbored acid-fast bacteria that reacted specifically with fluorescent antiserum to L. micdadei, confirmed by culture. In a patient at risk for nosocomial infection, the differential diagnosis of a positive sputum stain for acid-fast bacilli should include L. micdadei in addition to mycobacteria. Therapy for L. micdadei infection should be considered pending confirmation of the diagnosis. Images PMID:2430995

  13. Error-prone Replication Bypass of the Primary Aflatoxin B1 DNA Adduct, AFB1-N7-Gua*

    PubMed Central

    Lin, Ying-Chih; Li, Liang; Makarova, Alena V.; Burgers, Peter M.; Stone, Michael P.; Lloyd, R. Stephen

    2014-01-01

    Hepatocellular carcinomas (HCCs) are the third leading cause of cancer deaths worldwide. The highest rates of early onset HCCs occur in geographical regions with high aflatoxin B1 (AFB1) exposure, concomitant with hepatitis B infection. Although the carcinogenic basis of AFB1 has been ascribed to its mutagenic effects, the mutagenic property of the primary AFB1-DNA adduct, AFB1-N7-Gua, in mammalian cells has not been studied extensively. Taking advantage of the ability to create vectors containing a site-specific DNA adduct, the mutagenic potential was determined in primate cells. This adduct was highly mutagenic following replication in COS-7 cells, with a mutation frequency of 45%. The spectrum of mutations was predominantly G to T base substitutions, a result that is consistent with previous mutation data derived from aflatoxin-associated HCCs. To assess which DNA polymerases (pol) might contribute to the mutational outcome, in vitro replication studies were performed. Unexpectedly, replicative pol δ and the error-prone translesion synthesis pol ζ were able to accurately bypass AFB1-N7-Gua. In contrast, replication bypass using pol κ was shown to occur with low fidelity and could account for the commonly detected G to T transversions. PMID:24838242

  14. Aptamer induced assembly of fluorescent nitrogen-doped carbon dots on gold nanoparticles for sensitive detection of AFB1.

    PubMed

    Wang, Bin; Chen, Yanfen; Wu, Yuanya; Weng, Bo; Liu, Yingshuai; Lu, Zhisong; Li, Chang Ming; Yu, Cong

    2016-04-15

    Novel fluorescent nitrogen-doped carbon dots (N,C-dots) were synthesized and assembled on aptamer modified gold nanoparticles (Aptamer/AuNPs) for the super sensitive detection of aflatoxin B1 (AFB1). Positively charged N,C-dots were synthesized by the hydrothermal treatment of pancreatin. The prepared N,C-dots were assembled on aptamer/AuNPs by electrostatic interactions. The fluorescence of the N,C-dots was efficiently quenched. When AFB1 was added to the assay solution, specific interactions between AFB1 and the aptamer caused release of the N,C-dots. The fluorescence of the N,C-dots recovered and the intensity increase could be used to calculate the amount of AFB1 added. The assay exhibits super-high sensitivity with a detection limit of 5 pg/mL (16 pM) and a wide range of linear response of 5 pg/mL to 2.00 ng/mL. A novel aptasensor is thus successfully constructed, it provides an efficient way for sensitive AFB1 sensing as well as a new technique for aptamer based novel sensor construction. PMID:26584079

  15. Determination of the aflatoxin AFB1 from corn by direct analysis in real time-mass spectrometry (DART-MS).

    PubMed

    Busman, Mark; Liu, Jihong; Zhong, Hongjian; Bobell, John R; Maragos, Chris M

    2014-01-01

    Direct analysis in real time (DART) ionisation coupled to a high-resolution mass spectrometer (MS) was used for screening of aflatoxins from a variety of surfaces and the rapid quantitative analysis of a common form of aflatoxin, AFB1, extracted from corn. Sample preparation procedure and instrument parameter settings were optimised to obtain sensitive and accurate determination of aflatoxin AFB1. 84:16 acetonitrile water extracts of corn were analysed by DART-MS. The lowest calibration level (LCL) for aflatoxin AFB1 was 4 μg kg⁻¹. Quantitative analysis was performed with the use of matrix-matched standards employing the ¹³C-labelled internal standard for AFB1. DART-MS of spiked corn extracts gave linear response in the range 4-1000 μg kg⁻¹. Good recoveries (94-110%) and repeatabilities (RSD = 0.7-6.9%) were obtained at spiking levels of 20 and 100 μg kg⁻¹ with the use of an isotope dilution technique. Trueness of data obtained for AFB1 in maize by DART-MS was demonstrated by analysis of corn certified reference materials. PMID:24588621

  16. SCFTIR1/AFB-based auxin perception: mechanism and role in plant growth and development.

    PubMed

    Salehin, Mohammad; Bagchi, Rammyani; Estelle, Mark

    2015-01-01

    Auxin regulates a vast array of growth and developmental processes throughout the life cycle of plants. Auxin responses are highly context dependent and can involve changes in cell division, cell expansion, and cell fate. The complexity of the auxin response is illustrated by the recent finding that the auxin-responsive gene set differs significantly between different cell types in the root. Auxin regulation of transcription involves a core pathway consisting of the TIR1/AFB F-box proteins, the Aux/IAA transcriptional repressors, and the ARF transcription factors. Auxin is perceived by a transient coreceptor complex consisting of a TIR1/AFB protein and an Aux/IAA protein. Auxin binding to the coreceptor results in degradation of the Aux/IAAs and derepression of ARF-based transcription. Although the basic outlines of this pathway are now well established, it remains unclear how specificity of the pathway is conferred. However, recent results, focusing on the ways that these three families of proteins interact, are starting to provide important clues. PMID:25604443

  17. SCFTIR1/AFB-based auxin perception: mechanism and role in plant growth and development.

    PubMed

    Salehin, Mohammad; Bagchi, Rammyani; Estelle, Mark

    2015-01-01

    Auxin regulates a vast array of growth and developmental processes throughout the life cycle of plants. Auxin responses are highly context dependent and can involve changes in cell division, cell expansion, and cell fate. The complexity of the auxin response is illustrated by the recent finding that the auxin-responsive gene set differs significantly between different cell types in the root. Auxin regulation of transcription involves a core pathway consisting of the TIR1/AFB F-box proteins, the Aux/IAA transcriptional repressors, and the ARF transcription factors. Auxin is perceived by a transient coreceptor complex consisting of a TIR1/AFB protein and an Aux/IAA protein. Auxin binding to the coreceptor results in degradation of the Aux/IAAs and derepression of ARF-based transcription. Although the basic outlines of this pathway are now well established, it remains unclear how specificity of the pathway is conferred. However, recent results, focusing on the ways that these three families of proteins interact, are starting to provide important clues.

  18. SCFTIR1/AFB-Based Auxin Perception: Mechanism and Role in Plant Growth and Development

    PubMed Central

    Salehin, Mohammad; Bagchi, Rammyani; Estelle, Mark

    2015-01-01

    Auxin regulates a vast array of growth and developmental processes throughout the life cycle of plants. Auxin responses are highly context dependent and can involve changes in cell division, cell expansion, and cell fate. The complexity of the auxin response is illustrated by the recent finding that the auxin-responsive gene set differs significantly between different cell types in the root. Auxin regulation of transcription involves a core pathway consisting of the TIR1/AFB F-box proteins, the Aux/IAA transcriptional repressors, and the ARF transcription factors. Auxin is perceived by a transient coreceptor complex consisting of a TIR1/AFB protein and an Aux/IAA protein. Auxin binding to the coreceptor results in degradation of the Aux/IAAs and derepression of ARF-based transcription. Although the basic outlines of this pathway are now well established, it remains unclear how specificity of the pathway is conferred. However, recent results, focusing on the ways that these three families of proteins interact, are starting to provide important clues. PMID:25604443

  19. Normal probabilities for Vandenberg AFB wind components - monthly reference periods for all flight azimuths, 0- to 70-km altitudes

    NASA Technical Reports Server (NTRS)

    Falls, L. W.

    1975-01-01

    Vandenberg Air Force Base (AFB), California, wind component statistics are presented to be used for aerospace engineering applications that require component wind probabilities for various flight azimuths and selected altitudes. The normal (Gaussian) distribution is presented as a statistical model to represent component winds at Vandenberg AFB. Head tail, and crosswind components are tabulated for all flight azimuths for altitudes from 0 to 70 km by monthly reference periods. Wind components are given for 11 selected percentiles ranging from 0.135 percent to 99.865 percent for each month. The results of statistical goodness-of-fit tests are presented to verify the use of the Gaussian distribution as an adequate model to represent component winds at Vandenberg AFB.

  20. INDUCED SPUTUM DERIVES FROM THE CENTRAL AIRWAYS: CONFIRMATION USING A RADIOLABELED AEROSOL BOLUS DELIVERY TECHNIQUE

    EPA Science Inventory

    Indirect evidence suggests that induced sputum derives from the surfaces of the bronchial airways. To confirm this experimentally, we employed a radiolabeled aerosol bolus delivery technique that preferentially deposits aerosol in the central airways in humans. We hypothesized th...

  1. Utility of gram staining for evaluation of the quality of cystic fibrosis sputum samples.

    PubMed

    Nair, Bindu; Stapp, Jenny; Stapp, Lynn; Bugni, Linda; Van Dalfsen, Jill; Burns, Jane L

    2002-08-01

    The microscopic examination of Gram-stained sputum specimens is very helpful in the evaluation of patients with community-acquired pneumonia and has also been recommended for use in cystic fibrosis (CF) patients. This study was undertaken to evaluate that recommendation. One hundred one sputum samples from CF patients were cultured for gram-negative bacilli and examined by Gram staining for both sputum adequacy (using the quality [Q] score) and bacterial morphology. Subjective evaluation of adequacy was also performed and categorized. Based on Q score evaluation, 41% of the samples would have been rejected despite a subjective appearance of purulence. Only three of these rejected samples were culture negative for gram-negative CF pathogens. Correlation between culture results and quantitative Gram stain examination was also poor. These data suggest that subjective evaluation combined with comprehensive bacteriology is superior to Gram staining in identifying pathogens in CF sputum.

  2. Blood and sputum biomarkers in COPD and asthma: a review.

    PubMed

    Paone, G; Leone, V; Conti, V; De Marchis, L; Ialleni, E; Graziani, C; Salducci, M; Ramaccia, M; Munafò, G

    2016-01-01

    Chronic obstructive pulmonary disease (COPD) and asthma are lung inflammatory diseases that represent major public health problems. The primary, and often unique, method to evaluate lung function is spirometry, which reflects disease severity rather than disease activity. Moreover, its measurements strictly depend on patient's compliance, physician's expertise and data interpretation. The limitations of clinical history and pulmonary function tests have encouraged focusing on new possible tracers of diseases. The increase of the inflammatory response in the lungs represents an early pathological event, so biological markers related to inflammation may play key roles in earlier diagnosis, evaluation of functional impairment and prognosis. Biomarkers are measurable indicators associated with the presence and/or severity of a biological or pathogenic process, which may predict functional impairment, prognosis and response to therapy. The traditional approach based on invasive techniques (bronchoalveolar lavage and biopsies) may be replaced, at least in part, by using less invasive methods to collect specimens (sputum and blood), in which biomarkers could be measured. Proteomics, by the association between different protein profiles and pathogenic processes, is gaining an important role in pulmonary medicine allowing a more precise discrimination between patients with different outcomes and response to therapy. The aim of this review was to evaluate the use of biomarkers of airway inflammation in the context of both research and clinical practice. PMID:26957273

  3. Regular Sputum Check-Up for Early Diagnosis of Tuberculosis after Exposure in Healthcare Facilities

    PubMed Central

    Chao, Wen-Cheng; Wu, Chieh-Liang; Liu, Po-Yu; Shieh, Chi-Chang

    2016-01-01

    Background The early diagnosis of patients with TB disease is critical after an outbreak of tuberculosis (TB) infection in healthcare facilities. In this study, we report a catastrophic TB outbreak in a psychiatric healthcare facility and analyze the role of regular sputum check-ups and other diagnostic tools to facilitate an early diagnosis. Methods Every exposed participant received regular sputum check-ups and chest X-rays (CXR) as part of the outbreak management protocol. We retrospectively analyzed data from the contact participants to identify risk factors for eventual TB development and investigated the diagnostic efficacy of regular sputum check-ups. Results Among 133 contact participants, 16 (12.0%) developed TB during the 4-year follow-up period. Low body-mass-index (BMI) (<21) (adjusted hazard ratio (aHR) 3.16, 95% confidence interval (CI) 1.11–8.98) and long duration of contact (>3 months) (aHR 8.70, 95% CI, 1.14–63.34) independently predicted the development of TB. Even though regular sputum check-ups required significant resources, they did facilitate the early identification of new TB cases among the contact participants. Regular sputum check-ups for high-risk patients based on BMI, contact duration and CXR findings may be a practical approach when compared with universal sputum follow-up, with a slightly decreased sensitivity but high positive likelihood ratio (88%, [95% CI, 62–98%] and 5.12, [95%CI, 3.30–7.95], respectively). Conclusion While regular sputum check-ups for all contact participants facilitated the early identification of cases after the outbreak of TB in the healthcare facility, regular sputum check-ups for high-risk patients might be an effective alternative in resource-limited settings. PMID:27258370

  4. Altered Sputum Microstructure as a Marker of Airway Obstruction in Cystic Fibrosis Patients

    NASA Astrophysics Data System (ADS)

    Duncan, Gregg; Jung, James; West, Natalie; Boyle, Michael; Suk, Jung Soo; Hanes, Justin

    In the lungs of cystic fibrosis (CF) patients, highly viscoelastic mucus remains stagnant in the lung leading to obstructed airways prone to recurrent infections. Bulk-fluid rheological measurement is primarily used to assess the pathological features of mucus. However, this approach is limited in detecting microscopic properties on the length scale of pathogens and immune cells. We have shown in prior work based on the transport of muco-inert nanoparticles (MIP) in CF sputum that patients can carry significantly different microstructural properties. In this study, we aimed to determine the factors leading to variations between patients in sputum microstructure and their clinical implications. The microrheological properties of CF sputum were measured using multi-particle tracking experiments of MIP. MIP were made by grafting polyethylene glycol onto the surface of polystyrene nanoparticles which prior work has shown prevents adhesion to CF sputum. Biochemical analyses show that sputum microstructure was significantly altered by elevated mucin and DNA content. Reduction in sputum pore size is characteristic of patients with obstructed airways as indicated by measured pulmonary function tests. Our microstructural read-out may serve as a novel biomarker for CF.

  5. Evaluation of bleach-sedimentation for sterilising and concentrating Mycobacterium tuberculosis in sputum specimens

    PubMed Central

    2011-01-01

    Background Bleach-sedimentation may improve microscopy for diagnosing tuberculosis by sterilising sputum and concentrating Mycobacterium tuberculosis. We studied gravity bleach-sedimentation effects on safety, sensitivity, speed and reliability of smear-microscopy. Methods This blinded, controlled study used sputum specimens (n = 72) from tuberculosis patients. Bleach concentrations and exposure times required to sterilise sputum (n = 31) were determined. In the light of these results, the performance of 5 gravity bleach-sedimentation techniques that sterilise sputum specimens (n = 16) were compared. The best-performing of these bleach-sedimentation techniques involved adding 1 volume of 5% bleach to 1 volume of sputum, shaking for 10-minutes, diluting in 8 volumes distilled water and sedimenting overnight before microscopy. This technique was further evaluated by comparing numbers of visible acid-fast bacilli, slide-reading speed and reliability for triplicate smears before versus after bleach-sedimentation of sputum specimens (n = 25). Triplicate smears were made to increase precision and were stained using the Ziehl-Neelsen method. Results M. tuberculosis in sputum was successfully sterilised by adding equal volumes of 15% bleach for 1-minute, 6% for 5-minutes or 3% for 20-minutes. Bleach-sedimentation significantly decreased the number of acid-fast bacilli visualised compared with conventional smears (geometric mean of acid-fast bacilli per 100 microscopy fields 166, 95%CI 68-406, versus 346, 95%CI 139-862, respectively; p = 0.02). Bleach-sedimentation diluted paucibacillary specimens less than specimens with higher concentrations of visible acid-fast bacilli (p = 0.02). Smears made from bleach-sedimented sputum were read more rapidly than conventional smears (9.6 versus 11.2 minutes, respectively, p = 0.03). Counting conventional acid-fast bacilli had high reliability (inter-observer agreement, r = 0.991) that was significantly reduced (p = 0.03) by bleach

  6. Noise assessment of the rocket sled test track operation at Jolloman AFB, New Mexico. Final report

    SciTech Connect

    Shaffer, W.J.

    1988-10-01

    This report presents the results of noise data measurements of the Holloman AFB rocket-sled test-track operations. Impulse and community noise measurements were made to determine the impact of the rocket-sled noise on the surrounding community. A worst case sled run was measured and used to determine that the rocket sled has very little impact on the community for a worst-case rocket-sled run and little or no impact for the majority of the runs. Recommendations were made to limit the number of people exposed to the rocket sled noise and require test-track personnel to wear hearing protection. Sonic-boom measurement equipment should be purchased to document all sonic booms created by the rocket sled.

  7. Characterization of the geothermal resource at Lackland AFB, San Antonio, Texas. Phase I report

    SciTech Connect

    Lawford, T.W.; Malone, C.R.; Allman, D.W.; Zeisloft, J.; Foley, D.

    1983-06-01

    The geothermal resource under Lackland Air Force Base (AFB), San Antonio, Texas was studied. It is the conclusion of the investigators that a geothermal well drilled at the site recommended by this study has a high probability of delivering geothermal fluids in sufficient quantity and at adequate temperatures to support a projected space and domestic hot water heating system. An exploratory production well location is recommended in the southwest sector of the base, based upon geologic conditions and the availability of sufficient open space to support the drilling operation. It is projected that a production well drilled at the recommended location would produce geothermal fluid of 130 to 145/sup 0/F at a rate of approximately 1000 gpm with reasonable fluid drawdowns. The Environmental Assessment for the drilling portion of the project has been completed, and no irreversible or irretrievable impacts are anticipated as a result of this drilling program. The permitting process is proceeding smoothly.

  8. The lithostratigraphy of a marine kame delta-outwash fan complex at Pease AFB, Newington, NH

    SciTech Connect

    Dineen, R.J.; Manning, S.; McGeehan, K. )

    1993-03-01

    The overburden stratigraphy at Pease AFB is based on over 1,200 wells, borings, piezometers, and test pits, and includes five lithologic units: Fill, Upper Sand (US), Marine Clay and Silt (MCS), Lower Sand (LS), and Till (GT). The US is a yellow brown, poorly sorted sand to silty sand and is massive to laminated, and locally has hummocky bedding. The MCS (the Presumpscot Formation) is a dark gray, massive to laminated sandy to silty clay, and is locally interbedded with silty sand. The MCS contains a trace of organic matter, primarily as fine particles of peat. The LS is a gray to brown, poorly sorted, silty sand to gravelly sand that is massive to planar bedded and locally grades down into GT and/or upward into MCS. The GT consists of a massive to crudely bedded dark gray to dark brown, very poorly sorted, sandy silt to gravelly, silty sand. The US, MCS, LS and upper part of the GT were deposited in a marine environment at or near the ice margin. Pease AFB is built on two large fans of gravelly sand (LS plus US) that are bordered to the east by NW-SE till ridges (drumlins ). The northern-most fan is flat-topped with a surface elevation of 30 m ASL. The southern fan is more hummocky, with a surface elevation of 18.5 m ASL. Both fans coarsen towards the NW, and are interbedded with MCS towards the SE. The apices of the fans overlie deeply-scoured troughs in the rock surface. The fans are interpreted to be kame deltas or submarine outwash fans that are deposited along the retreating Wisconsinan ice margin by concentrated meltwater flow. Later, the US may have been deposited by marine shoreface erosion of the emergent fans as the ice front retreated and sea level fell.

  9. 33 CFR 334.1130 - Pacific Ocean, Western Space and Missile Center (WSMC), Vandenberg AFB, Calif.; danger zones.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 33 Navigation and Navigable Waters 3 2012-07-01 2012-07-01 false Pacific Ocean, Western Space and... RESTRICTED AREA REGULATIONS § 334.1130 Pacific Ocean, Western Space and Missile Center (WSMC), Vandenberg AFB, Calif.; danger zones. (a) The Area. (1) The waters of the Pacific Ocean in an area extending...

  10. 33 CFR 334.1130 - Pacific Ocean, Western Space and Missile Center (WSMC), Vandenberg AFB, Calif.; danger zones.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 3 2011-07-01 2011-07-01 false Pacific Ocean, Western Space and... RESTRICTED AREA REGULATIONS § 334.1130 Pacific Ocean, Western Space and Missile Center (WSMC), Vandenberg AFB, Calif.; danger zones. (a) The Area. (1) The waters of the Pacific Ocean in an area extending...

  11. 33 CFR 334.1130 - Pacific Ocean, Western Space and Missile Center (WSMC), Vandenberg AFB, Calif.; danger zones.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 33 Navigation and Navigable Waters 3 2013-07-01 2013-07-01 false Pacific Ocean, Western Space and... RESTRICTED AREA REGULATIONS § 334.1130 Pacific Ocean, Western Space and Missile Center (WSMC), Vandenberg AFB, Calif.; danger zones. (a) The Area. (1) The waters of the Pacific Ocean in an area extending...

  12. 33 CFR 334.1130 - Pacific Ocean, Western Space and Missile Center (WSMC), Vandenberg AFB, Calif.; danger zones.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Pacific Ocean, Western Space and... RESTRICTED AREA REGULATIONS § 334.1130 Pacific Ocean, Western Space and Missile Center (WSMC), Vandenberg AFB, Calif.; danger zones. (a) The Area. (1) The waters of the Pacific Ocean in an area extending...

  13. Appendix C -- Soil gas analysis of vadose zone monitoring system installation report for McClellan AFB

    SciTech Connect

    1996-10-31

    This appendix compiles the data from gas chromatography/mass spectroscopy measurements of gas samples from McClellan AFB soils collected and analyzed in December, 1995. Compounds detected include: vinyl chloride, methylene chloride, chloroethanes, chloroethenes, benzene, chlorobenzenes, propylene, trimethylbenzenes, acetone, toluene, xylenes.

  14. Determination of the aflatoxin AFB1 from corn by direct analysis in real time-mass spectrometry (DART-MS)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Direct analysis in real time (DART) ionization coupled to a high resolution mass spectrometer (MS) was used for screening of aflatoxins from a variety of surfaces and the rapid quantitative analysis of a common form of aflatoxin, AFB1, extracted from corn. Sample preparation procedure and instrument...

  15. 33 CFR 334.1130 - Pacific Ocean, Western Space and Missile Center (WSMC), Vandenberg AFB, Calif.; danger zones.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 33 Navigation and Navigable Waters 3 2014-07-01 2014-07-01 false Pacific Ocean, Western Space and... RESTRICTED AREA REGULATIONS § 334.1130 Pacific Ocean, Western Space and Missile Center (WSMC), Vandenberg AFB, Calif.; danger zones. (a) The area. (1) The waters of the Pacific Ocean in an area extending...

  16. The Most Common Detected Bacteria in Sputum of Patients with the Acute Exacerbation of COPD

    PubMed Central

    Cukic, Vesna

    2013-01-01

    Introduction: Acute exacerbation of COPD (AECOPD) may be triggered by infection with bacteria or viruses or by environmental pollutants; the cause of about one-third of exacerbations cannot be identified. Objective: To determine the most common bacteria in sputum culture of patients with AECOPD hospitalized in Intensive care unit of Clinic for pulmonary disease and TB “Podhrastovi” in the 2012. Material and methods: This is a retrospective analysis of sputum bacterial cultures of patients with AECOPD treated in the Intensive care unit of Clinic for pulmonary disease and TB “Podhrastovi” during 2012 .year. Each patient was required to give two sputum for bacterial examination. Each patient was treated with antibiotics prior to admission in Clinic “Podhrastovi”. The results of sputum bacterial culture findings are expressed in absolute number and percentage of examined patients. Results: In 2012, 75 patients with AECOPD were treated in Intensive care unit of Clinic for pulmonary disease and TB“Podhrastovi”. 44 (58.66%) of patients had normal –nonpathogenic – usual bacterial flora isolated in sputum cultures, 31 (41.34%) had a pathogen bacteria in sputum culture as follows: 7 had Streptoccocus pneumoniae, 8 had Klebsiella pneumoniae (2 with Streptococcus pneumoniae, one with Acinetobacter baumani) ,4 Escherichia colli, others are one or two cases with other bacteria. Conclusion: Bacterial airway infections play a great role in many, but not in all, of cases of AECOPD. So there is the need to do a sputum bacterial culture examination in each patient with AECOPD and with appropriate antibiotics to contribute to curing of them. PMID:24511262

  17. Throat Swabs and Sputum Culture as Predictors of P. aeruginosa or S. aureus Lung Colonization in Adult Cystic Fibrosis Patients

    PubMed Central

    Seidler, Darius; Griffin, Mary; Nymon, Amanda; Koeppen, Katja; Ashare, Alix

    2016-01-01

    Background Due to frequent infections in cystic fibrosis (CF) patients, repeated respiratory cultures are obtained to inform treatment. When patients are unable to expectorate sputum, clinicians obtain throat swabs as a surrogate for lower respiratory cultures. There is no clear data in adult subjects demonstrating the adequacy of throat swabs as a surrogate for sputum or BAL. Our study was designed to determine the utility of throat swabs in identifying lung colonization with common organisms in adults with CF. Methods Adult CF subjects (n = 20) underwent bronchoscopy with BAL. Prior to bronchoscopy, a throat swab was obtained. A sputum sample was obtained from subjects who were able to spontaneously expectorate. All samples were sent for standard microbiology culture. Results Using BAL as the gold standard, we found the positive predictive value for Pseudomonas aeruginosa to be 100% in both sputum and throat swab compared to BAL. However, the negative predictive value for P. aeruginosa was 60% and 50% in sputum and throat swab, respectively. Conversely, the positive predictive value for Staphylococcus aureus was 57% in sputum and only 41% in throat swab and the negative predictive value of S. aureus was 100% in sputum and throat swab compared to BAL. Conclusions Our data show that positive sputum and throat culture findings of P. aeruginosa reflect results found on BAL fluid analysis, suggesting these are reasonable surrogates to determine lung colonization with P. aeruginosa. However, sputum and throat culture findings of S. aureus do not appear to reflect S. aureus colonization of the lung. PMID:27711152

  18. Semi-Automated, Occupationally Safe Immunofluorescence Microtip Sensor for Rapid Detection of Mycobacterium Cells in Sputum

    PubMed Central

    Soelberg, Scott D.; Weigel, Kris M.; Hiraiwa, Morgan; Cairns, Andrew; Lee, Hyun-Boo; Furlong, Clement E.; Oh, Kieseok; Lee, Kyong-Hoon; Gao, Dayong; Chung, Jae-Hyun; Cangelosi, Gerard A.

    2014-01-01

    An occupationally safe (biosafe) sputum liquefaction protocol was developed for use with a semi-automated antibody-based microtip immunofluorescence sensor. The protocol effectively liquefied sputum and inactivated microorganisms including Mycobacterium tuberculosis, while preserving the antibody-binding activity of Mycobacterium cell surface antigens. Sputum was treated with a synergistic chemical-thermal protocol that included moderate concentrations of NaOH and detergent at 60°C for 5 to 10 min. Samples spiked with M. tuberculosis complex cells showed approximately 106-fold inactivation of the pathogen after treatment. Antibody binding was retained post-treatment, as determined by analysis with a microtip immunosensor. The sensor correctly distinguished between Mycobacterium species and other cell types naturally present in biosafe-treated sputum, with a detection limit of 100 CFU/mL for M. tuberculosis, in a 30-minute sample-to-result process. The microtip device was also semi-automated and shown to be compatible with low-cost, LED-powered fluorescence microscopy. The device and biosafe sputum liquefaction method opens the door to rapid detection of tuberculosis in settings with limited laboratory infrastructure. PMID:24465845

  19. Significant roadblocks exist in developing sputum sample libraries for clinical validation of novel in vitro diagnostics

    PubMed Central

    Dollow, Joshua M; Green, Justin A

    2014-01-01

    With the continuing rise of multiresistant pathogens, reliable, cost-effective, and novel diagnostics are urgently required by clinicians and clinical trialists to diagnose conditions such as respiratory tract infections to enable rational antimicrobial choice and enhance clinical outcomes. However, during product development, validation of these in vitro diagnostic devices, a key regulatory hurdle, requires sputum samples in large numbers. The Rapid Point-of-Care test Platform for Infectious Diseases (RAPP-ID) consortium is tasked with producing point of care test (POCT) platforms for rapid diagnosis of lower respiratory tract infections, including tuberculosis and blood stream infections. Validation of diagnostic platforms would ideally use well-characterized samples in a sputum library taken from a range of clinical settings to allow for a wide panel of pathogens to be assessed. These samples would be stored in specific stable conditions (monitored temperature, specific medium) until required for validation. Therefore we reviewed the current literature for details of storage conditions of sputum samples and for previous validation studies of other diagnostic tests using this methodology. However, we conclude that little data exists, and thus the acquisition and successful storage of good quality clinical samples are major roadblocks in the validation of novel POCT platforms, and that while not without limitations, spiked sputum samples appear the best solution until sputum library laboratory techniques allowing careful preservation of pathogens are improved. PMID:24489460

  20. Clinical and immunological characteristics associated with the presence of protozoa in sputum smears.

    PubMed

    Martínez-Girón, Rafael; van Woerden, Hugo Cornelis

    2013-01-01

    The objective of this study is to assess the relationship between protozoa in spontaneously expectorated sputum samples and a range of clinical and immunological variables. Clinical details including age, gender, smoking status, and use of oral or inhaled steroids were recorded for a cohort of 199 patients whose spontaneously expectorated sputum samples were submitted to a Cytology Laboratory in Spain between January 2005 and December 2006. Slides were scanned for protozoa under light microscopy and scanned for monocytes/small macrophages highlighted by immunocytochemistry (CD68 monoclonal antibody). One hundred ninety-one patients provided adequate sputum samples, of whom 70 had protozoa in their sputum. There was a strong relationship between the presence of protozoa and monocytes/small macrophages identified under light microscopy (P < 0.001). A binary logistic regression model also indicated a relationship between protozoa and both smoking status and steroid use. The diagnoses in those with protozoa included infection (including tuberculosis), chronic obstructive pulmonary disease (COPD), lung fibrosis, asthma, chronic liver disease, immunosuppression, cancer, pancreatic or renal disease, heart failure, and AIDS. The identified association between protozoa and monocytes/small macrophages in sputum suggests an immune response and warrants further investigation to clarify whether or not these organisms have any pathological significance in this wide range of conditions.

  1. Sputum eosinophils from asthmatics express ICAM-1 and HLA-DR.

    PubMed Central

    Hansel, T T; Braunstein, J B; Walker, C; Blaser, K; Bruijnzeel, P L; Virchow, J C; Virchow, C

    1991-01-01

    Sputum from symptomatic asthmatics is a rich source of eosinophils from the respiratory tract. Following liquefaction of sputum with dithioerythritol (DTE), a cell suspension for indirect double immunofluorescence with flow cytometry was obtained. Eosinophils were identified using anti-CD9 fluorescein conjugate, and particular surface markers measured with the relevant mouse MoAb followed by goat anti-mouse immunoglobulin phycoerythrin conjugate. Blood and sputum eosinophil surface markers were determined in parallel from asthmatics not receiving steroid therapy. Sputum eosinophils were found to have considerably elevated levels of CD11b, a reflection of eosinophil activation. Sputum but not blood eosinophils were found to express ICAM-1 (nine out of 11 cases) and HLA-DR (eight out of 11 cases). Furthermore, following culture of normal blood eosinophils with pooled T cell supernatants, ICAM-1 and HLA-DR could be induced in vitro. The induction of eosinophil adhesion molecules such as ICAM-1 and HLA-DR may influence eosinophil localization and function in asthma. PMID:1682072

  2. Semi-automated, occupationally safe immunofluorescence microtip sensor for rapid detection of Mycobacterium cells in sputum.

    PubMed

    Inoue, Shinnosuke; Becker, Annie L; Kim, Jong-Hoon; Shu, Zhiquan; Soelberg, Scott D; Weigel, Kris M; Hiraiwa, Morgan; Cairns, Andrew; Lee, Hyun-Boo; Furlong, Clement E; Oh, Kieseok; Lee, Kyong-Hoon; Gao, Dayong; Chung, Jae-Hyun; Cangelosi, Gerard A

    2014-01-01

    An occupationally safe (biosafe) sputum liquefaction protocol was developed for use with a semi-automated antibody-based microtip immunofluorescence sensor. The protocol effectively liquefied sputum and inactivated microorganisms including Mycobacterium tuberculosis, while preserving the antibody-binding activity of Mycobacterium cell surface antigens. Sputum was treated with a synergistic chemical-thermal protocol that included moderate concentrations of NaOH and detergent at 60°C for 5 to 10 min. Samples spiked with M. tuberculosis complex cells showed approximately 10(6)-fold inactivation of the pathogen after treatment. Antibody binding was retained post-treatment, as determined by analysis with a microtip immunosensor. The sensor correctly distinguished between Mycobacterium species and other cell types naturally present in biosafe-treated sputum, with a detection limit of 100 CFU/mL for M. tuberculosis, in a 30-minute sample-to-result process. The microtip device was also semi-automated and shown to be compatible with low-cost, LED-powered fluorescence microscopy. The device and biosafe sputum liquefaction method opens the door to rapid detection of tuberculosis in settings with limited laboratory infrastructure.

  3. Decontamination of sputum for longer time in sodium hydroxide for isolation of Mycobacterium tuberculosis.

    PubMed

    Satapathy, P; Das, D; Murmu, B N; Kar, S K

    2014-12-01

    Decontamination by modified Petroff's method is being practiced in many laboratories carrying out Mycobacterium tuberculosis culture and drug susceptibility testing. The method exposes mycobacteria to 4% sodium hydroxide for 30min. However, laboratories in developing countries with limited resources might be using a type of centrifuge that does not open during power failures and exposes the mycobacteria to alkali for longer periods. Out of 28 smear-positive specimens processed, 85.7%, 85.7% and 60.7% of specimens showed a positive culture after exposure to alkali for 0.5, 1.0 and 72h. Laboratories compelled to expose the mycobacteria for a longer duration of time can still attempt isolation for culture as only a small amount of bacteria are needed for culture positivity. PMID:26786630

  4. Immuno-physiological alterations from AFB1 in rats counteracted by treatments with Lactobacillus paracasei BEJ01 and montmorillonite clay mixture.

    PubMed

    Ben Salah-Abbès, Jalila; Jebali, Rania; Sharafi, Hakimeh; Akbari Noghabi, Kambiz; Oueslati, Ridha; Abbès, Samir

    2016-09-01

    High contamination by aflatoxin B1 (AFB1) has been detected in Beja province (Tunisia) in many dairy products and animal feed, which has resulted in many tons of cereals and cereals being removed from the market, causing economic loss. While removal represents a means of reducing risk, exposures still occur. Studies have increasingly focused on means of AFB1 biodegradation/elimination using lactic acid bacteria and clay mineral. In the study here, Lactobacillus paracasei BEJ01 (LP) and montmorilonite clay (MT) were used to reduce the physio-/immunotoxicologic disorders that could develop in rats that underwent AFB1 exposures for a total of 7 consecutive days. The results indicated that rats treated with AFB1 (80 μg/kg BW) alone had significant decreases in lymphocytes in their blood (including B-lymphocytes, CD3(+), CD4(+), and CD8(+) T-lymphocyte subtypes, and NK cells), immunoglobulins (IgA and IgG) and pro-inflammatory cytokines; these rats also had altered oxidative stress status. In contrast, in rats treated with LP + MT (2 × 10(9) cfu/ml [∼ 2 mg/kg] + 0.5 mg MT/kg BW) for a total of 7 days before, concurrent with or after AFB1 treatment, there was a significant blockade/mitigation of each AFB1-impacted parameter. Moreover, treatment with the mixture at any point in relation to AFB1 treatment expectedly caused enhanced TNFα and IL-1β expression relative to control values; all other parameters were comparable to values noted in control rats. Alone, the mixture had no impact on host parameters. From the results here it may be concluded the the LP + MT mixture was effective in protecting these hosts against AFB1-induced immunologic/physiologic disorders and that LP + MT could prevent and/or mitigate AFB1 toxicities in vivo.

  5. Immuno-physiological alterations from AFB1 in rats counteracted by treatments with Lactobacillus paracasei BEJ01 and montmorillonite clay mixture.

    PubMed

    Ben Salah-Abbès, Jalila; Jebali, Rania; Sharafi, Hakimeh; Akbari Noghabi, Kambiz; Oueslati, Ridha; Abbès, Samir

    2016-09-01

    High contamination by aflatoxin B1 (AFB1) has been detected in Beja province (Tunisia) in many dairy products and animal feed, which has resulted in many tons of cereals and cereals being removed from the market, causing economic loss. While removal represents a means of reducing risk, exposures still occur. Studies have increasingly focused on means of AFB1 biodegradation/elimination using lactic acid bacteria and clay mineral. In the study here, Lactobacillus paracasei BEJ01 (LP) and montmorilonite clay (MT) were used to reduce the physio-/immunotoxicologic disorders that could develop in rats that underwent AFB1 exposures for a total of 7 consecutive days. The results indicated that rats treated with AFB1 (80 μg/kg BW) alone had significant decreases in lymphocytes in their blood (including B-lymphocytes, CD3(+), CD4(+), and CD8(+) T-lymphocyte subtypes, and NK cells), immunoglobulins (IgA and IgG) and pro-inflammatory cytokines; these rats also had altered oxidative stress status. In contrast, in rats treated with LP + MT (2 × 10(9) cfu/ml [∼ 2 mg/kg] + 0.5 mg MT/kg BW) for a total of 7 days before, concurrent with or after AFB1 treatment, there was a significant blockade/mitigation of each AFB1-impacted parameter. Moreover, treatment with the mixture at any point in relation to AFB1 treatment expectedly caused enhanced TNFα and IL-1β expression relative to control values; all other parameters were comparable to values noted in control rats. Alone, the mixture had no impact on host parameters. From the results here it may be concluded the the LP + MT mixture was effective in protecting these hosts against AFB1-induced immunologic/physiologic disorders and that LP + MT could prevent and/or mitigate AFB1 toxicities in vivo. PMID:27294391

  6. Lung Microbiome Analysis in Steroid-Naїve Asthma Patients by Using Whole Sputum

    PubMed Central

    Jung, Jae-Woo; Choi, Jae-Chol; Shin, Jong-Wook; Kim, Jae-Yeol; Park, In-Won; Choi, Byoung Whui; Park, Heung-Woo; Cho, Sang-Heon

    2016-01-01

    Background Although recent metagenomic approaches have characterized the distinguished microbial compositions in airways of asthmatics, these results did not reach a consensus due to the small sample size, non-standardization of specimens and medication status. We conducted a metagenomics approach by using terminal restriction fragment length polymorphism (T-RFLP) analysis of the induced whole sputum representing both the cellular and fluid phases in a relative large number of steroid naïve asthmatics. Methods Induced whole sputum samples obtained from 36 healthy subjects and 89 steroid-naїve asthma patients were analyzed through T-RFLP analysis. Results In contrast to previous reports about microbiota in the asthmatic airways, the diversity of microbial composition was not significantly different between the controls and asthma patients (p=0.937). In an analysis of similarities, the global R-value showed a statistically significant difference but a very low separation (0.148, p=0.002). The dissimilarity in the bacterial communities between groups was 28.74%, and operational taxonomic units (OTUs) contributing to this difference were as follows: OTU 789 (Lachnospiraceae), 517 (Comamonadaceae, Acetobacteraceae , and Chloroplast), 633 (Prevotella), 645 (Actinobacteria and Propionibacterium acnes), 607 (Lactobacillus buchneri, Lactobacillus otakiensis, Lactobacillus sunkii, and Rhodobacteraceae), and 661 (Acinetobacter, Pseudomonas, and Leptotrichiaceae), and they were significantly more prevalent in the sputum of asthma patients than in the sputum of the controls. Conclusion Before starting anti-asthmatic treatment, the microbiota in the whole sputum of patients with asthma showed a marginal difference from the microbiota in the whole sputum of the controls. PMID:27433177

  7. A novel flow cytometric-based method to measure kinase inhibition in sputum from COPD subjects

    PubMed Central

    Nicholson, G C; Holloway, R A; Leaker, B R; Kilty, I; Salganik, M; Tan, L; Barnes, P J; Donnelly, L E

    2016-01-01

    Introduction Janus kinases (JAKs) regulate inflammatory gene expression through phosphorylation of signal transducer and activator of transcription (STAT) proteins. Expression of STAT proteins is increased in chronic obstructive pulmonary disease (COPD), and may be involved in driving chronic inflammation. Oral JAK inhibitors are effective as anti-inflammatory therapy but exhibit dose-limiting adverse effects. Development of inhaled compounds would be enhanced by robust biomarkers that directly reflect the anti-inflammatory and pharmacological activity in the lung. Methods A novel flow cytometry assay was developed to measure STAT1 phosphorylation in sputum inflammatory cells. The standard sputum processing method was refined to improve sputum cell viability. The flow cytometric assay was used to assess the reproducibility of the measurement of STAT1 phosphorylation and the in vitro activity of a pan JAK-inhibitor on three separate visits in patients with COPD. Results Upregulation of STAT1 phosphorylation was measured following in vitro IFNγ stimulation of sputum macrophages (stimulated/unstimulated ratio 1.57; p<0.00001). Upregulation was inhibited following in vitro preincubation with a pan JAK-inhibitor (inhibited+stimulated/unstimulated ratio 0.97). STAT1 phosphorylation activity could only be measured in macrophages. Conclusions Sputum from patients with COPD can be used to reproducibly measure phospho-STAT expression in sputum macrophages. The flow cytometry-based method can be used to evaluate kinase inhibitors in vitro and subsequently in ex vivo studies. The assay is particularly useful for the assessment of inhaled compounds where whole blood assays may not be relevant. PMID:27403320

  8. Novel methodology to assess sputum smear microscopy quality in private laboratories

    PubMed Central

    2012-01-01

    Background In South Asia, it is estimated that 80% of patients choose to attend a private facility for their healthcare needs. Although patients generally believe that the private-sector provides high quality services, private diagnostic laboratories are largely unregulated and little is known about the accuracy of results provided. This study assesses the accuracy of sputum smear microscopy for pulmonary tuberculosis diagnosis in private laboratories operating in Karachi, Pakistan. A novel evaluation methodology was designed in which patient-actors submitted sputum specimens spiked with cultured Mycobacterium tuberculosis (Mtb) for testing such that laboratories were not aware that they were being assessed. Methods Smear-negative sputum specimens from Indus Hospital TB Program patients were collected and combined with an attenuated, cultured Mtb strain to create Mtb-spiked samples; for negative standards, no Mtb was added to the smear-negative sputum specimens. Seven of the largest private laboratories across Karachi were chosen for evaluation and were sent six Mtb-spiked and one Mtb-negative sputum specimens. Patient-actors pretending to be laboratory customers submitted these specimens to each laboratory for testing over a three day period. Results Only three laboratories accurately classified all the Mtb-spiked specimens which were submitted. A further three misclassified all the Mtb-spiked specimens as smear-negative, thus providing the ‘patients’ with false negative results. Conclusions TB sputum smear microscopy services are highly variable across private laboratories and are often of extremely poor quality. Engagement, capacity building and rigorous monitoring of standards at private laboratories are of vital importance for the control of TB. Our findings, while specific for TB diagnostic tests, could be symptomatic of other tests performed in private laboratories and warrant further investigation. PMID:23193964

  9. Investigating the quality of expectorated sputum for tuberculosis diagnosis in Bolivia.

    PubMed

    Hernández, P; Punchak, M; Camacho, M; Hepple, P; McNerney, R

    2015-09-01

    A low-power microscope-based cytological system to assess the quality of expectorated sputum provided for tuberculosis (TB) diagnosis was piloted in Bolivia. A total of 3688 samples were subjected to visual and cytological examination in nine laboratories: of these, 591 (16%) were misclassified by visual examination and 294 (8%) were found to be degraded. The degree of discordance varied between locations, and laboratories received a higher number of degraded specimens from isolated health clinics. Cytological assessment of sputum was found to be feasible and identified areas for improvement in the Bolivian diagnostic system for TB.

  10. A method for double-labeling sputum cells for p53 and cytokeratin

    SciTech Connect

    Neft, R.E.; Tierney, L.A.; Belinsky, S.A.

    1995-12-01

    Molecular and immunological techniques may enhance the usefulness of sputum cytology as a screening tool for lung cancer. These techniques may also be useful in detecting and following the early progression of disease from metaplasia to dysplasia, carcinoma in situ, and finally to invasive carcinoma. Longitudinal information on the evolution of these malignant changes in the respiratory epithelium can be gained by prospective study of populations at high risk for lung cancer. This work is significant because double-labeling of cells in sputum with p53 and cytokeratin antibodies facilitates rapid screening of p53 positive neoplastic and preneoplastic lung cells by brightfield and fluorescence microscopy.

  11. Fate of sputum samples transported in bottles containing cetylpyridinium chloride and sodium chloride: a national reference laboratory study.

    PubMed

    Visalakshi, Potharaju; Meharwal, Sandeep K; Arora, Jyoti; Bhalla, Manpreet; Singh, Niti; Myneedu, Vithal Prasad; Behera, Digamber

    2012-04-01

    The transportation of sputum samples may sometimes take more than one week which results in an increased contamination rate and loss of positive cultures. The current study was planned to analyze the recovery rate of mycobacteria from transported samples with and without Cetylpyridinium chloride (CPC). Addition of CPC is useful for isolation of M. tuberculosis from sputum subjected to long-term storage.

  12. The penetration of fresh undiluted sputum expectorated by cystic fibrosis patients by non-adhesive polymer nanoparticles

    PubMed Central

    Suk, Jung Soo; Lai, Samuel K.; Wang, Ying-Ying; Ensign, Laura M.; Zeitlin, Pamela L.; Boyle, Michael P.; Hanes, Justin

    2009-01-01

    Highly viscoelastic and adhesive sputum has precluded efficient nanoparticle-based drug and gene delivery to the lungs of patients with cystic fibrosis (CF). We sought to determine whether nanoparticles coated with non-mucoadhesive polymers could penetrate CF sputum, and to use these “muco-inert particles” (MIPs) as non-destructive nanoprobes to characterize the sputum microstructure. Particles as large as 200 nm in diameter that were densely coated with low MW polyethylene glycol (PEG) moved through undiluted CF sputum with average speeds up to 90-fold faster than similarly-sized uncoated particles. On the other hand, the transport of both coated and uncoated 500 nm particles was strongly hindered. The local viscosity of sputum, encountered by the fastest 10% of 200 nm MIPs, was only 5-fold higher than that of water, whereas the bulk viscosity was 10,000-fold higher at low shear rates. Using measured transport rates of various sized MIPs combined with an obstruction-scaling model, we determined that the average 3D mesh spacing of CF sputum is ~140 ± 50 nm (range: 60-300 nm). Taken together, these results demonstrate that nanoparticles up to 200 nm in diameter that do not adhere to CF sputum can move rapidly through this critical barrier by accessing pores that are filled with a low viscosity fluid. The results also offer hope that desperately needed sputum-penetrating drug- and gene-carrier nanoparticles can be developed for CF. PMID:19176245

  13. Vadose zone monitoring system installation report for McClellan AFB

    SciTech Connect

    Zawislanski, P.; Faybishenko, B.; James, A.; Freifeld, B.; Salve, R.

    1996-10-31

    Two vadose zone monitoring systems (VZMS) have been installed at Site S-7, in Investigation Cluster 34 (IC 34), in Operable Unit A (OU A) of McClellan AFB. The two boreholes, VZMS-A and VZMS-B were instrumented at depths ranging from approximately 6 ft to 113 ft. Instruments were installed in clusters using a custom-made stainless steel cage with a spring-loaded mechanism allowing instruments to be in contact with the well bore wall once in place. Each cluster contains a tensiometer, suction lysimeter, soil gas probe and thermistor for measuring hydraulic potential, liquid- and gas-phase pressure, temperature of the formation and for collecting samples for chemical analyses in both the liquid and gas phases. Neutron probe logging is performed in two separate, smaller borings, VZMS-NP-1 and VZMS-NP-2, to obtain soil moisture content data. Preliminary details of soil gas analyses, laboratory field testing of soil samples, particle size analyses and neutron probe data are presented.

  14. Shemya AFB, Alaska 1992 IRP field investigation report. Volume 2, Appendix A: Final report

    SciTech Connect

    Not Available

    1993-02-01

    The US Air Force is currently investigating 22 sites on Shemya Air Force Base (AFB) to determine if past spill and disposal activities have caused environmental damage. These investigations are being carried out under the Air Force`s Installation Restoration Program (IRP). Field investigations were performed in 1992 to obtain the information needed to assess what future actions will need to be carried out at each site. The island`s drinking water supply was also investigated. Activities completed at 10 selected sites during the 1992 field investigation included surface sampling to determine the lateral extent of contamination, subsurface sampling to determine the vertical extent of contamination, and the installation of well points and monitoring wells to determine the direction of groundwater flow and if the groundwater has been affected by a site. In addition, geophysical surveys were performed at most sites to identify site boundaries and check for the presence of buried metal to be avoided during drilling activities. This report, appendix A, contains the analytical results.

  15. Shemya AFB, Alaska 1992 IRP field investigation report. Volume 1: Final report

    SciTech Connect

    Not Available

    1993-02-01

    The US Air Force is currently investigating 22 sites on Shemya Air Force Base (AFB) to determine if past spill and disposal activities have caused environmental damage. These investigations are being carried out under the Air Force`s Installation Restoration Program (IRP). As a part of the IRP program, field investigations were performed in 1992 to obtain the information needed to assess what future actions willneed to be carried out at each site. The island`s drinking water supply was also investigated. Activities completed at 10 selected sites during the 1992 field investigation included surface sampling to determine the lateral extent of contamination, subsurface sampling to determine the vertical extent of contamination, and the installation of well points and monitoring wells to determine the direction of groundwater flow and if the groundwater has been affected by a site. In addition, geophysical surveys were performed at most sites to identify site boundaries and check for the presence of buried metal to be avoided during drilling activities.

  16. Shemya AFB, Alaska 1992 IRP field investigation report. Volume 4, Appendixes E and F: Final report

    SciTech Connect

    Not Available

    1993-02-01

    The US Air Force is currently investigating 22 sites on Shemya Air Force Base (AFB) to determine if past spill and disposal activities have caused environmental damage. These investigations are being carried out under the Air Force`s Installation Restoration Program (IRP). Field investigations were performed in 1992 to obtain the information needed to assess what future actions will need to be carried out at each site. The island`s drinking water supply was also investigated. Activities completed at 10 selected sites included surface sampling to determine the lateral extent of contamination, subsurface sampling to determine the vertical extent of contamination, and the installation of well points and monitoring wells to determine the direction of groundwater flow and if the groundwater has been affected by a site. In addition, geophysical surveys were performed at most sites to identify site boundaries and check for the presence of buried metal, to be avoided during drilling activities. This report contains appendices E and F with information on the following: soil boring logs, and data validation of samples analyzed.

  17. Wastewater characterization survey, Victor Valley Wastewater Reclamation Authority and hazardous-waste survey at George AFB, California. Final report

    SciTech Connect

    Binovi, R.D.; Ng, E.K.; Tetla, R.A.

    1987-01-01

    This is a report of a survey of the Victor Wastewater Reclamation Authority Sewerage system, the sewage treatment plant, and effluent from the various operations at George AFB, California. The scope of work included the characterization of the wastewater from George AFB, as well as characterization of effluents from 29 oil/water separators servicing industrial operations on base, flow measurements at three locations on base, a microbiological evaluation of aeration basin foam, bench-scale activated-sludge studies, and a review of results from previous surveys. Recommendations: (1) AFFF (Aqueous Film Forming Foam) should never be discharged to the sewer. (2) Programming for pretreatment should proceed at selected operations. (3) More waste and wastestream analysis be performed. (4) Upgrade waste accumulation points. (5) Implement an aggressive inspection program for oil/water separators. (6) Cut down on nonessential washing.

  18. Ultra-rapid real-time PCR for the detection of Paenibacillus larvae, the causative agent of American Foulbrood (AFB).

    PubMed

    Han, Sang-Hoon; Lee, Do-Bu; Lee, Dong-Woo; Kim, Eul-Hwan; Yoon, Byoung-Su

    2008-09-01

    A novel micro-PCR-based detection method, termed ultra-rapid real-time PCR, was applied to the development of a rapid detection for Paenibacillus larvae (P. larvae) which is the causative agent of American Foulbrood (AFB). This method was designed to detect the 16S rRNA gene of P. larvae with a micro-scale chip-based real-time PCR system, GenSpector TMC-1000, which has uncommonly fast heating and cooling rates (10 degrees C per second) and small reaction volume (6microl). In the application of ultra-rapid real-time PCR detection to an AFB-infected larva, the minimum detection time was 7 min and 54s total reaction time (30 cycles), including the melting temperature analysis. To the best of our knowledge, this novel detection method is one of the most rapid real-time PCR-based detection tools. PMID:18571197

  19. Calcareous concretions and psammoma bodies in sputum smears: do these similar structures have different clinical significance?

    PubMed

    Martínez-Girón, Rafael; Martínez-Torre, Santiago; Tamargo-Peláez, María Luisa; López-Cabanilles, María Dolores; Torre-Bayón, Concepción

    2014-09-01

    Different noncellular elements, such as round concentric calcified laminated structures, may be found in sputum smears. If these structures appear isolated on the background of the smear, the term usually used to describe them is "calcareous concretions" (CC). On the contrary, when the structures are part of epithelial cell groups or small tissue fragments, the term used to describe them is "Psammoma bodies" (PB). The aim of this work is to establish the relationship between these structures and pulmonary disease, especially lung carcinoma, by searching for the presence of CC and/or PB in sputum smears. Our study has taken as a basis 16.716 sputum smears from 696 patients obtained during a 7-year period (2003-2009). After reviewing them, it was found that from the total, 66 cases (0.39%) contained round calcified structures, 57 of them (0.34%) corresponding to CC, and the remaining 9 ones (0.05%) corresponding to PB. From these 57 CC cases, 56 corresponded to benign entities, and only one was found with lung carcinoma. On the other hand, from the 9 PB cases all of them (100%) were related to lung adenocarcinoma. We conclude that, even having a similar morphological structure, these aforementioned calcified structures we have observed in sputum smears have different and relevant clinical significance.

  20. Sputum Leucine-Rich Alpha-2 Glycoprotein as a Marker of Airway Inflammation in Asthma

    PubMed Central

    Honda, Hiromi; Fujimoto, Minoru; Miyamoto, Shintaro; Ishikawa, Nobuhisa; Serada, Satoshi; Hattori, Noboru; Nomura, Shintaro; Kohno, Nobuoki; Yokoyama, Akihito; Naka, Tetsuji

    2016-01-01

    Background Asthma is a chronic inflammatory disease of airways, but an ideal biomarker that accurately reflects ongoing airway inflammation has not yet been established. The aim of this study was to examine the potential of sputum leucine-rich alpha-2 glycoprotein (LRG) as a new biomarker for airway inflammation in asthma. Methods We obtained induced sputum samples from patients with asthma (N = 64) and healthy volunteers (N = 22) and measured LRG concentration by sandwich enzyme-linked immunosorbent assay (ELISA). Ovalbumin (OVA)-induced asthma model mice were used to investigate the mechanism of LRG production during airway inflammation. The LRG concentrations in the bronchoalveolar lavage fluid (BALF) obtained from mice were determined by ELISA and mouse lung sections were stained with anti-LRG antibody and periodic acid-Schiff (PAS) reagent. Results Sputum LRG concentrations were significantly higher in patients with asthma than in healthy volunteers (p = 0.00686). Consistent with patients’ data, BALF LRG levels in asthma model mice were significantly higher than in control mice (p = 0.00013). Immunohistochemistry of lung sections from asthma model mice revealed that LRG was intensely expressed in a subpopulation of bronchial epithelial cells, which corresponded with PAS-positive mucus producing cells. Conclusion These findings suggest that sputum LRG is a promising biomarker of local inflammation in asthma. PMID:27611322

  1. A SENSITIVE IMMUNOFLUORESCENCE ASSAY FOR DETECTION OF P53 PROTEIN ACCUMULATION IN SPUTUM

    EPA Science Inventory

    p53 mutations are common genetic alterations in lung cancers and usually result in p53 protein accumulation in tumor cells. Sputum is noninvasive to collect and ideal for screening p53 abnormalities. This study was to determine the feasibility of detecting p53 protein accumulatio...

  2. In vivo effects of recombinant human DNase I on sputum in patients with cystic fibrosis.

    PubMed Central

    Shah, P. L.; Scott, S. F.; Knight, R. A.; Marriott, C.; Ranasinha, C.; Hodson, M. E.

    1996-01-01

    BACKGROUND: Viscoelastic secretions in cystic fibrosis cause impaired mucus clearance and persistence of bacteria within the lung. The abnormal rheology is partly due to the presence of high molecular weight deoxyribonucleic acid (DNA). Recombinant human DNase I (rhDNase) has been shown to depolymerise DNA and thereby reduce the in vitro viscoelasticity of sputum in patients with cystic fibrosis. A phase II double blind placebo controlled study showed that rhDNase improved pulmonary function in patients with cystic fibrosis. The object of the present study was to evaluate the in vivo effects of rhDNase on sputum rheology and to determine whether these were correlated with changes in pulmonary function. METHODS: Patients were randomised to receive either placebo or rhDNase 2.5 mg twice daily for 10 days. Sputum samples were collected in sterile containers during screening and during treatment with the study drug. Pulmonary function and rheological analysis were the primary outcomes evaluated. Other parameters assessed were quantitative sputum bacteriology, sputum DNA concentration, and change in molecular mass of DNA polymers. RESULTS: The viscoelasticity of the sputum in untreated patients with cystic fibrosis was high and treatment with rhDNase reduced all the rheological parameters measured: dynamic storage modulus (a measure of elasticity), dynamic loss modulus (a measure of viscosity), and log complex modulus (a measure of mucus rigidity). The calculated cough clearance index was also improved following treatment with rhDNase. These rheological parameters showed a correlation with forced expiratory volume in one second (FEV1) which was improved by a mean (SE) of 13.3 (5.6)% on day 10 of treatment with rhDNase compared with a change of 0.2 (3.1)% in the placebo group. There was no change in bacterial colony counts or sputum DNA concentrations following treatment with rhDNase, but a small decrease in high molecular weight DNA was observed. CONCLUSIONS: Patients

  3. Detection of serum AFB1-lysine adduct in Malaysia and its association with liver and kidney functions.

    PubMed

    Mohd Redzwan, S; Rosita, Jamaluddin; Mohd Sokhini, A M; Nurul 'Aqilah, A R; Wang, Jia-Sheng; Kang, Min-Su; Zuraini, Ahmad

    2014-01-01

    Aflatoxin is ubiquitously found in many foodstuffs and produced by Aspergillus species of fungi. Of many aflatoxin metabolites, AFB1 is classified by the International Agency for Research on Cancer (IARC) as group one carcinogen and linked to the development of hepatocellular carcinoma (HCC). The study on molecular biomarker of aflatoxin provides a better assessment on the extent of human exposure to aflatoxin. In Malaysia, the occurrences of aflatoxin-contaminated foods have been documented, but there is a lack of data on human exposure to aflatoxin. Hence, this study investigated the occurrence of AFB1-lysine adduct in serum samples and its association with liver and kidney functions. 5ml fasting blood samples were collected from seventy-one subjects (n=71) for the measurement of AFB1-lysine adduct, albumin, total bilirubin, AST (aspartate aminotransferase), ALT (alanine transaminase), ALP (alkaline phosphatase), GGT (gamma-glutamyl transpeptidase), creatinine and BUN (blood urea nitrogen). The AFB1-lysine adduct was detected in all serum samples (100% detection rate) with a mean of 6.85±3.20pg/mg albumin (range: 1.13-18.85pg/mg albumin). Male subjects (mean: 8.03±3.41pg/mg albumin) had significantly higher adduct levels than female subjects (mean: 5.64±2.46pg/mg albumin) (p<0.01). It was noteworthy that subjects with adduct levels greater than average (>6.85pg/mg albumin) had significantly elevated level of total bilirubin (p<0.01), GGT (p<0.05) and creatinine (p<0.01). Nevertheless, only the level of total bilirubin, (r=0.347, p-value=0.003) and creatinine (r=0.318, p-value=0.007) showed significant and positive correlation with the level of AFB1-lysine adduct. This study provides a valuable insight on human exposure to aflatoxin in Malaysia. Given that aflatoxin can pose serious problem to the health, intervention strategies should be implemented to limit/reduce human exposure to aflatoxin. Besides, a study with a big sample size should be warranted in

  4. Aberrant promoter methylation of multiple genes in sputum from individuals exposed to smoky coal emissions

    PubMed Central

    Liu, Yang; Lan, Qing; Shen, Min; Mumford, Judy; Keohavong, Phouthone

    2010-01-01

    Summary Aberrant methylation in the promoter region of cancer-related genes leads to gene transcriptional inactivation and plays an integral role in lung tumorigenesis. Recent studies demonstrated that promoter methylation was detected not only in lung tumors from patients with lung cancer but also in sputum of smokers without the disease, suggesting the potential for aberrant gene promoter methylation in sputum as a predictive marker for lung cancer. In the present study, we investigated promoter methylation of 4 genes frequently detected in lung tumors, including p16, MGMT, RASSF1A and DAPK genes, in sputum samples obtained from 107 individuals, including 34 never-smoking females and 73 mostly smoking males, who had no evidence of lung cancer but who were exposed to smoky coal emission in Xuan Wei County, China, where lung cancer rate is more than 6 times the Chinese national average rate. Forty nine of the individuals showed evidence of chronic bronchitis while the remaining 58 individuals showed no such a symptom. Promoter methylation of p16, MGMT, RASSF1A and DAPK was detected in 51.4% (55/107), 17.8% (19/107), 29.9% (32/107), and 15.9% (17/107) of the sputum samples from these individuals, respectively. There were no differences in promoter methylation frequencies of any of these genes according to smoking status or gender of the subjects or between individuals with chronic bronchitis and those without evidence of such a symptom. Therefore, individuals exposed to smoky coal emissions in this region harbored in their sputum frequent promoter methylation of these genes that have been previously found in lung tumors and implicated in lung cancer development. PMID:18751376

  5. Ventilator versus manual hyperinflation in clearing sputum in ventilated intensive care unit patients.

    PubMed

    Dennis, Diane; Jacob, Wendy; Budgeon, Charley

    2012-01-01

    The aim of hyperinflation in the ventilated intensive care unit patient is to increase oxygenation, reverse lung collapse and clear sputum. The efficacy and consistency of manual hyperventilation is well supported in the literature, but there is limited published evidence supporting hyperventilation utilising a ventilator. Despite this, a recent survey established that almost 40% of Australian tertiary intensive care units utilise ventilator hyperinflation. The aim of this non-inferiority cross-over study was to determine whether ventilator hyperinflation was as effective as manual hyperinflation in clearing sputum from patients receiving mechanical ventilation using a prescriptive ventilator hyperinflation protocol. Forty-six patients received two randomly ordered physiotherapy treatments on the same day by the same physiotherapist. The efficacy of the hyperinflation modes was measured by sputum wet weight. Secondary measures included compliance, tidal volume, airway pressure and PaO2/FiO2 ratio. There was no difference in wet weight of sputum cleared using ventilator hyperinflation or manual hyperinflation (mean 3.2 g, P=0.989). Further, no difference in compliance (P=0.823), tidal volume (P=0.219), heart rate (P=0.579), respiratory rate (P=0.929) or mean arterial pressure (P=0.593) was detected. A statistically significant difference was seen in mean airway pressure (P=0.002) between techniques. The effect of techniques on the PaO2/FiO2 response ratio was dependent on time (interaction P=0.024). Physiotherapy using ventilator hyperinflation cleared a comparable amount of sputum and was as safe as manual hyperinflation. This research describes a ventilator hyperinflation protocol that will serve as a platform for continued discussion, research and development of its application in ventilated patients.

  6. Microbial, host and xenobiotic diversity in the cystic fibrosis sputum metabolome

    PubMed Central

    Quinn, Robert A; Phelan, Vanessa V; Whiteson, Katrine L; Garg, Neha; Bailey, Barbara A; Lim, Yan Wei; Conrad, Douglas J; Dorrestein, Pieter C; Rohwer, Forest L

    2016-01-01

    Cystic fibrosis (CF) lungs are filled with thick mucus that obstructs airways and facilitates chronic infections. Pseudomonas aeruginosa is a significant pathogen of this disease that produces a variety of toxic small molecules. We used molecular networking-based metabolomics to investigate the chemistry of CF sputa and assess how the microbial molecules detected reflect the microbiome and clinical culture history of the patients. Metabolites detected included xenobiotics, P. aeruginosa specialized metabolites and host sphingolipids. The clinical culture and microbiome profiles did not correspond to the detection of P. aeruginosa metabolites in the same samples. The P. aeruginosa molecules that were detected in sputum did not match those from laboratory cultures. The pseudomonas quinolone signal (PQS) was readily detectable from cultured strains, but absent from sputum, even when its precursor molecules were present. The lack of PQS production in vivo is potentially due to the chemical nature of the CF lung environment, indicating that culture-based studies of this pathogen may not explain its behavior in the lung. The most differentially abundant molecules between CF and non-CF sputum were sphingolipids, including sphingomyelins, ceramides and lactosylceramide. As these highly abundant molecules contain the inflammatory mediator ceramide, they may have a significant role in CF hyperinflammation. This study demonstrates that the chemical makeup of CF sputum is a complex milieu of microbial, host and xenobiotic molecules. Detection of a bacterium by clinical culturing and 16S rRNA gene profiling do not necessarily reflect the active production of metabolites from that bacterium in a sputum sample. PMID:26623545

  7. Evaluation of bromide mass discharge in a sandy aquifer at Vandenberg AFB, CA

    NASA Astrophysics Data System (ADS)

    Mackay, D. M.; Rasa, E.; Einarson, M.; Kaiser, P.; Chakraborty, I.; Scow, K. M.

    2009-12-01

    Side-by-side experiments were conducted by UC Davis research team at a former fuel station at Vandenberg Air Force Base (AFB) to evaluate the rate of transformation of methyl tert-butyl ether (MTBE) to tert-butyl alcohol (TBA) impacted by ethanol and to investigate evidence of TBA degradation under sulfate reducing conditions. On one side we injected groundwater amended with ethanol and MTBE. In the other lane we injected groundwater amended with TBA. On both sides, injected ground water was spiked with bromide tracer to provide estimates of groundwater flow direction variations, flow velocity, dispersion, and mobile mass loss resulting from diffusive sequestration into aquitards. 162 monitoring wells were aligned into seven transects located downgradient of the injection wells. The mass discharge approach was used to evaluate the natural attenuation of the injected constituents. In this talk we will focus on calculations of mass discharge of the bromide tracer at each of the seven monitoring well transects. The amount of bromide mass discharged through each transect was calculated for any sampling time using field measurements of break through curves. Cumulative mass discharges were estimated and, by iteration based on mass balance, the flow properties of the aquifer were estimated. The calibration process resulted in subtle but quantitatively important changes in our assumptions regarding key physical properties of the aquifer (thickness, porosity) which could be only approximately estimated by standard methods (coring, CPT, etc.). On the basis of this calibration, a more robust approach was devised for evaluating the source and fate of TBA in the aquifer.

  8. Remediation of NAPL source zones: lessons learned from field studies at Hill and Dover AFB.

    PubMed

    McCray, John E; Tick, Geoffrey R; Jawitz, James W; Gierke, John S; Brusseau, Mark L; Falta, Ronald W; Knox, Robert C; Sabatini, David A; Annable, Michael D; Harwell, Jeffrey H; Wood, A Lynn

    2011-01-01

    Innovative remediation studies were conducted between 1994 and 2004 at sites contaminated by nonaqueous phase liquids (NAPLs) at Hill and Dover AFB, and included technologies that mobilize, solubilize, and volatilize NAPL: air sparging (AS), surfactant flushing, cosolvent flooding, and flushing with a complexing-sugar solution. The experiments proved that aggressive remedial efforts tailored to the contaminant can remove more than 90% of the NAPL-phase contaminant mass. Site-characterization methods were tested as part of these field efforts, including partitioning tracer tests, biotracer tests, and mass-flux measurements. A significant reduction in the groundwater contaminant mass flux was achieved despite incomplete removal of the source. The effectiveness of soil, groundwater, and tracer based characterization methods may be site and technology specific. Employing multiple methods can improve characterization. The studies elucidated the importance of small-scale heterogeneities on remediation effectiveness, and fomented research on enhanced-delivery methods. Most contaminant removal occurs in hydraulically accessible zones, and complete removal is limited by contaminant mass stored in inaccessible zones. These studies illustrated the importance of understanding the fluid dynamics and interfacial behavior of injected fluids on remediation design and implementation. The importance of understanding the dynamics of NAPL-mixture dissolution and removal was highlighted. The results from these studies helped researchers better understand what processes and scales are most important to include in mathematical models used for design and data analysis. Finally, the work at these sites emphasized the importance and feasibility of recycling and reusing chemical agents, and enabled the implementation and success of follow-on full-scale efforts.

  9. Mannitol versus hypertonic saline: Safety and efficacy of mannitol and hypertonic saline in sputum induction and bronchial hyperreactivity assessment.

    PubMed

    Alvarez-Puebla, M J; Olaguibel, J M; Almudevar, E; Echegoyen, A A; Vela, C; de Esteban, B

    2015-08-01

    Eosinophilic asthma phenotype predicts good response to corticosteroids and associates to asthmatic exacerbations. Sputum induction by hypertonic saline (HS) inhalation is technically demanding. Bronchial hyperresponsiveness (BHR) to osmotic agents indirectly mirrors active airway inflammation. We compared the safety and ability of HS and mannitol to induce sputum and measure BHR. We evaluated the stability of inflammatory phenotypes. We studied 35 non-smoking asthmatics randomized to undergo HS and mannitol challenges on 2 days 1 week apart. Sputum was sampled for cell analysis and phenotyped as eosinophilic (≥3% eosinophils) and non-eosinophilic (<3%) asthma. Nineteen subjects had BHR to mannitol and nine of them also had BHR to HS. Drops in forced expiratory volume in 1 s were higher from HS challenge than from mannitol challenge. Adequate sputum samples were obtained from 80% subjects (68% mannitol and 71% HS). Eosinophils and macrophages from both challenges correlated. Neutrophils were higher in sputum from HS. Ninety percent samples were equally phenotyped with HS and mannitol. Fractional exhaled nitric oxide, sputum eosinophils and BHR correlated in both challenges. HS and mannitol showed similar capacity to produce valuable sputum samples. BHR to both osmotic stimuli partially resembled airway eosinophilic inflammation but mannitol was more sensitive than HS to assess BHR. Eosinophilic phenotype remained stable in most patients with both stimuli.

  10. Smoking status and anti-inflammatory macrophages in bronchoalveolar lavage and induced sputum in COPD

    PubMed Central

    2011-01-01

    Background Macrophages have been implicated in the pathogenesis of COPD. M1 and M2 macrophages constitute subpopulations displaying pro- and anti-inflammatory properties. We hypothesized that smoking cessation affects macrophage heterogeneity in the lung of patients with COPD. Our aim was to study macrophage heterogeneity using the M2-marker CD163 and selected pro- and anti-inflammatory mediators in bronchoalveolar lavage (BAL) fluid and induced sputum from current smokers and ex-smokers with COPD. Methods 114 COPD patients (72 current smokers; 42 ex-smokers, median smoking cessation 3.5 years) were studied cross-sectionally and underwent sputum induction (M/F 99/15, age 62 ± 8 [mean ± SD] years, 42 (31-55) [median (range)] packyears, post-bronchodilator FEV1 63 ± 9% predicted, no steroids past 6 months). BAL was collected from 71 patients. CD163+ macrophages were quantified in BAL and sputum cytospins. Pro- and anti-inflammatory mediators were measured in BAL and sputum supernatants. Results Ex-smokers with COPD had a higher percentage, but lower number of CD163+ macrophages in BAL than current smokers (83.5% and 68.0%, p = 0.04; 5.6 and 20.1 ×104/ml, p = 0.001 respectively). The percentage CD163+ M2 macrophages was higher in BAL compared to sputum (74.0% and 30.3%, p < 0.001). BAL M-CSF levels were higher in smokers than ex-smokers (571 pg/ml and 150 pg/ml, p = 0.001) and correlated with the number of CD163+ BAL macrophages (Rs = 0.38, p = 0.003). No significant differences were found between smokers and ex-smokers in the levels of pro-inflammatory (IL-6 and IL-8), and anti-inflammatory (elafin, and Secretory Leukocyte Protease Inhibitor [SLPI]) mediators in BAL and sputum. Conclusions Our data suggest that smoking cessation partially changes the macrophage polarization in vivo in the periphery of the lung towards an anti-inflammatory phenotype, which is not accompanied by a decrease in inflammatory parameters. PMID:21426578

  11. Calcium montmorillonite clay reduces AFB1 and FB1 biomarkers in rats exposed to single and co-exposures of aflatoxin and fumonisin

    PubMed Central

    Mitchell, Nicole J.; Xue, Kathy S.; Lin, Shuhan; Marroquin-Cardona, Alicia; Brown, Kristal A.; Elmore, Sarah E.; Tang, Lili; Romoser, Amelia; Gelderblom, Wentzel C. A.; Wang, Jia-Sheng; Phillips, Timothy D.

    2014-01-01

    Aflatoxins (AFs) and fumonisins (FBs) can co-contaminate foodstuffs and have been associated with hepatocellular and esophageal carcinomas in humans at high risk for exposure. One strategy to reduce exposure (and toxicity) from contaminated foodstuffs is the dietary inclusion of a montmorillonite clay (UPSN) that binds AFs and FBs in the GI tract. In this study, the binding capacity of UPSN was evaluated for AFB1, FB1 and a combination thereof in Fischer-344 rats. Rats were pre-treated with different dietary levels of UPSN (0.25 or 2%) for 1 week. Rats were gavaged with a single dose of either 0.125 mg AFB1 or 25 mg FB1/kg b.w. and a combination thereof in the presence and absence of an aqueous solution of UPSN. The kinetics of mycotoxin excretion were monitored by analyzing serum AFB1-albumin, urinary AF (AFM1), and FB1 biomarkers over a period of 72 hr. UPSN decreased AFM1 excretion by 88-97%, indicating highly effective binding. FB1 excretion was reduced, to a lesser extent, ranging between 45 to 85%. When in combination, both AFB1 and FB1 binding occurred, but capacity was decreased by almost half. In the absence of UPSN, the combined AFB1 and FB1 treatment decreased the urinary biomarkers by 67 and 45% respectively, but increased levels of AFB1-albumin, presumably by modulating its cytochrome metabolism. UPSN significantly reduced bioavailability of both AFB1 and FB1 when in combination; suggesting that it can be utilized to reduce levels below their respective thresholds for affecting adverse biological effects. PMID:24193864

  12. Calcium montmorillonite clay reduces AFB1 and FB1 biomarkers in rats exposed to single and co-exposures of aflatoxin and fumonisin.

    PubMed

    Mitchell, Nicole J; Xue, Kathy S; Lin, Shuhan; Marroquin-Cardona, Alicia; Brown, Kristal A; Elmore, Sarah E; Tang, Lili; Romoser, Amelia; Gelderblom, Wentzel C A; Wang, Jia-Sheng; Phillips, Timothy D

    2014-07-01

    Aflatoxins (AFs) and fumonisins (FBs) can co-contaminate foodstuffs and have been associated with hepatocellular and esophageal carcinomas in humans at high risk for exposure. One strategy to reduce exposure (and toxicity) from contaminated foodstuffs is the dietary inclusion of a montmorillonite clay (UPSN) that binds AFs and FBs in the gastrointestinal tract. In this study, the binding capacity of UPSN was evaluated for AFB1, FB1 and a combination thereof in Fischer 344 rats. Rats were pre-treated with different dietary levels of UPSN (0.25% or 2%) for 1 week. Rats were gavaged with a single dose of either 0.125 mg AFB1 or 25 mg FB1 per kg body weight and a combination thereof in the presence and absence of an aqueous solution of UPSN. The kinetics of mycotoxin excretion were monitored by analyzing serum AFB1 -albumin, urinary AF (AFM1) and FB1 biomarkers over a period of 72 h. UPSN decreased AFM1 excretion by 88-97%, indicating highly effective binding. FB1 excretion was reduced, to a lesser extent, ranging from 45% to 85%. When in combination, both AFB1 and FB1 binding occurred, but capacity was decreased by almost half. In the absence of UPSN, the combined AFB1 and FB1 treatment decreased the urinary biomarkers by 67% and 45% respectively, but increased levels of AFB1 -albumin, presumably by modulating its cytochrome metabolism. UPSN significantly reduced bioavailability of both AFB1 and FB1 when in combination; suggesting that it can be utilized to reduce levels below their respective thresholds for affecting adverse biological effects. PMID:24193864

  13. Defining Binding Efficiency and Specificity of Auxins for SCFTIR1/AFB-Aux/IAA Co-receptor Complex Formation

    PubMed Central

    2013-01-01

    Structure–activity profiles for the phytohormone auxin have been collected for over 70 years, and a number of synthetic auxins are used in agriculture. Auxin classification schemes and binding models followed from understanding auxin structures. However, all of the data came from whole plant bioassays, meaning the output was the integral of many different processes. The discovery of Transport Inhibitor-Response 1 (TIR1) and the Auxin F-Box (AFB) proteins as sites of auxin perception and the role of auxin as molecular glue in the assembly of co-receptor complexes has allowed the development of a definitive quantitative structure–activity relationship for TIR1 and AFB5. Factorial analysis of binding activities offered two uncorrelated factors associated with binding efficiency and binding selectivity. The six maximum-likelihood estimators of Efficiency are changes in the overlap matrixes, inferring that Efficiency is related to the volume of the electronic system. Using the subset of compounds that bound strongly, chemometric analyses based on quantum chemical calculations and similarity and self-similarity indices yielded three classes of Specificity that relate to differential binding. Specificity may not be defined by any one specific atom or position and is influenced by coulomb matrixes, suggesting that it is driven by electrostatic forces. These analyses give the first receptor-specific classification of auxins and indicate that AFB5 is the preferred site for a number of auxinic herbicides by allowing interactions with analogues having van der Waals surfaces larger than that of indole-3-acetic acid. The quality factors are also examined in terms of long-standing models for the mechanism of auxin binding. PMID:24313839

  14. Monitoring and data analysis for the Vadose Zone Monitoring System (VZMS), McClellan AFB. Quarterly status report

    SciTech Connect

    Zawislanski, P.T.; Salve, R.; Freifeld, B.

    1997-05-28

    This report contains information on field and laboratory work performed between January and May 15th 1997 at site S-7 in IC 34, at McClellan AFB. At this location, a Vadose Zone Monitoring System (VZMS) is currently being used to collect subsurface data including hydraulic potential, soil gas pressure, moisture content, water chemistry, gas chemistry, and temperature. Due to delays in the completion of the above-ground installations, data collection did not commence until mid-February. As a result, the data presented in this report is preliminary.

  15. Sputum screening for lung cancer in radon exposed uranium miners: a comparison of semi-automated sputum cytometry and conventional cytology.

    PubMed

    Marek, W; Richartz, G; Philippou, S; Marek, L; Kotschy-Lang, N

    2007-11-01

    Preparing for a prospective study on early lung cancer, correlation between semi-automated sputum cytometry (ASC) and conventional cytology (CY) was examined in 1517 former uranium miners with posterior-anterior and lateral chest roentgenograms. A hundred and twenty sputum specimens were classified as suspicious (grade II) and another 18 as highly suspicious (grade III) by ASC. Within grade III group, 9 samples were classified by CY as tumor cell positive, 7 severe, and 1 mild and 1 moderate dysplasias. In the group of grade II ASC, 7 were tumor cell positive, 27 classified as severe dysplasia or CIS, 20 as moderate and 19 as mild dysplasia. Twenty seven contained metaplasias and 18 were normal or inflammatory. Of the 1358 samples classified as benign (grade I) by ASC, only 5 samples were classified by CY as severe dysplasia, 6 as moderate and 34 as mild dysplasia, 173 as metaplasia, the others were normal or inflammatory. Twenty one samples were judged as inadequate for ASC and CY. At present, 23 tumors were found in final diagnosis. Sensitivity of ASC was 87% at a specificity of 92%, while CY, at high grade alterations as a threshold, had a sensitivity of 83% at 97% specificity. We conclude that, along with modern radiological procedures and molecular biological markers, ASC and CY should be included in a controlled prospective randomized study on early lung cancer.

  16. [Drug resistance testing of Mycobacterium tuberculosis isolates from sputum in Chad].

    PubMed

    Abdelhadi, O; Ndokaïn, J; Ali, M Moussa; Friocourt, V; Mortier, E; Heym, B

    2012-02-01

    Culture and resistance testing of Mycobacterium tuberculosis are not regularly performed in Chad. Sputa were obtained from three different categories of hospitals (district, regional and national) in Chad. All examined sputa were smear-positive and were investigated by culture and drug resistance testing for first-line antituberculosis drugs. From 232 sputa positive for acid-fast bacilli, 135 isolates of M. tuberculosis from different patients (46 women, 89 men, mean age 34 years) were analyzed. All the patients except one corresponded to new cases of tuberculosis. In total, 27 out of 135 isolates (20%) were resistant to at least one major antituberculosis drug. Resistance to isoniazid was the most frequent resistance observed, with 18 isolates (13%) presenting at least this resistance. Three isolates (2.2%) were resistant to isoniazid and rifampicin (multidrug resistance MDR) including one isolate being concomitantly resistant to streptomycin and ethambutol. The resistance rate differed in relation to the category of the hospital; the most important resistance rate was observed in regional hospitals (33%), while it was 16% and 14% in the national and district hospitals, respectively. HIV serology was performed in 81 patients, among whom 20 (25%) were positive. This is the first study that shows that drug resistance of M. tuberculosis is present in Chad. Besides single drug-resistant isolates, multidrug-resistant strains of M. tuberculosis could also be identified. This result highlights the urgency of initiating actions to detect drug resistance and limit the spread of drug-resistant strains.

  17. Recombinant Human DNase I Reduces the Viscosity of Cystic Fibrosis Sputum

    NASA Astrophysics Data System (ADS)

    Shak, Steven; Capon, Daniel J.; Hellmiss, Renate; Marsters, Scot A.; Baker, Carrie L.

    1990-12-01

    Respiratory distress and progressive lung destruction in cystic fibrosis can be attributed to bacterial persistence and the accumulation of viscous purulent secretions in the airways. More than 30 yr ago it was suggested that the large amounts of DNA in purulent secretions contribute to its viscosity and that bovine pancreatic DNase I could reduce the viscosity. To evaluate the potential clinical utility of recombinant human DNase I (rhDNase) in the treatment of cystic fibrosis, we have cloned, sequenced, and expressed rhDNase. Catalytic amounts of rhDNase greatly reduce the viscosity of purulent cystic fibrosis sputum, transforming it within minutes from a nonflowing viscous gel to a flowing liquid. The reduction in viscosity is associated with a decrease in size of DNA in the sputum. Inhalation of a rhDNase aerosol may be a simple direct approach that will help individuals with cystic fibrosis and other patients with pneumonia or bronchitis to clear their airways of purulent secretions.

  18. Effect of corticosteroids on sputum sol-phase protease inhibitors in chronic obstructive pulmonary disease.

    PubMed Central

    Wiggins, J; Elliott, J A; Stevenson, R D; Stockley, R A

    1982-01-01

    Corticosteroids caused a reduction in the ratio of sol-phase sputum concentration to serum concentration of albumin in 12 patients with chronic obstructive bronchitis, suggesting a reduction in protein transudation. Alpha-1-antitrypsin values followed the same pattern as those of albumin in both the control and treatment periods, confirming the similar behaviour of the two proteins. The alpha 1-antichymotrypsin ratios were on average three times higher than those of albumin in the control period, confirming the presence of local mechanisms in the lung for preferentially concentrating this protein. The sputum-to-serum ratio of alpha 1-antichymotrypsin, however, rose during steroid treatment with the result that there was a selective increase in this protease inhibitor, which may be of potential benefit to such patients. PMID:6984237

  19. Smoking Cessation and the Microbiome in Induced Sputum Samples from Cigarette Smoking Asthma Patients

    PubMed Central

    Munck, Christian; Helby, Jens; Westergaard, Christian G.; Porsbjerg, Celeste; Backer, Vibeke; Hansen, Lars H.

    2016-01-01

    Asthma is a common disease causing cough, wheezing and shortness of breath. It has been shown that the lung microbiota in asthma patients is different from the lung microbiota in healthy controls suggesting that a connection between asthma and the lung microbiome exists. Individuals with asthma who are also tobacco smokers experience more severe asthma symptoms and smoking cessation is associated with improved asthma control. In the present study we investigated if smoking cessation in asthma patients is associated with a change in the bacterial community in the lungs, examined using induced sputum. We found that while tobacco smokers with asthma have a greater bacterial diversity in the induced sputum compared to non-smoking healthy controls, smoking cessation does not lead to a change in the microbial diversity. PMID:27391160

  20. Bacterial flora in the sputum and comorbidity in patients with acute exacerbations of COPD

    PubMed Central

    Boixeda, Ramon; Almagro, Pere; Díez-Manglano, Jesús; Cabrera, Francisco Javier; Recio, Jesús; Martin-Garrido, Isabel; Soriano, Joan B

    2015-01-01

    Objective To determine in patients admitted with an acute exacerbation of chronic obstructive pulmonary disease (AE-COPD) the association between the isolation of potential pathogens in a conventional sputum culture and comorbidities. Patients and methods The ESMI study is a multicenter observational study. Patients with AE-COPD admitted to the Internal Medicine departments of 70 hospitals were included. The clinical characteristics, treatments, and comorbidities were gathered. The results of conventional sputum cultures were recorded. Results A total of 536 patients were included, of which 161 produced valid sputum and a potentially pathogenic microorganism was isolated from 88 subjects (16.4%). The isolation of Pseudomonas aeruginosa (30.7%) was associated with a greater severity of the lung disease (previous admissions [P= 0.026], dyspnea scale [P=0.047], post-broncodilator forced expiratory volume in 1 second (FEV1) [P=0.005], and the BODEx index [P=0.009]); also with higher prevalence of cor pulmonale (P=0.017), heart failure (P=0.048), and cerebrovascular disease (P=0.026). Streptococcus pneumoniae (26.1%) was associated with more comorbidity according to number of diseases (P=0.018); notably, peripheral artery disease (P=0.033), hypertension (P=0.029), dyslipidemia (P=0.039), osteoporosis (P=0.0001), and depression (P=0.005). Conclusion Patients with AE-COPD and P. aeruginosa present higher severity of COPD, while those with S. pneumoniae present greater comorbidity. The potentially pathogenic microorganism obtained in the sputum culture depends on the associated comorbidities. PMID:26664106

  1. Clinical and Radiographic Manifestations of Sputum Culture-Negative Pulmonary Tuberculosis

    PubMed Central

    Nguyen, Minh-Vu H.; Jenny-Avital, Elizabeth R.; Burger, Susanne; Leibert, Eric M.; Achkar, Jacqueline M.

    2015-01-01

    Intervention at the earliest possible stage of pulmonary tuberculosis (PTB) reduces morbidity for the individual and transmission for the community. We characterize the clinical and radiographic manifestations of sputum culture-negative (Cx-) PTB in order to facilitate awareness of this under recognized and likely early disease state. In this cross-sectional sub-study, we reviewed the medical records of HIV-uninfected PTB patients enrolled from 2006–2014 within the context of a TB biomarker study in New York City. Cx- PTB was defined as clinical and/or radiographic presentation consistent with PTB, three initial mycobacterial sputum cultures negative, and no evidence of other respiratory disease. Diagnosis was confirmed by clinical and radiographic improvement on antituberculous treatment and/or culture, nucleic acid, or histological confirmation from a specimen other than the initial three sputa. Cx+ PTB was defined as above but with M. tuberculosis growth in at least one of the first three sputum cultures. Demographics, symptoms, and radiographic findings on initial presentation were compared between the two groups. Of 99 subjects diagnosed with PTB, 21 met the criteria of Cx- PTB. Cx- compared to Cx+ subjects presented with a significantly lower frequency of cough (70% vs. 91%, P = 0.02), sputum production (30% vs. 64%, P < 0.01), weight loss (25% vs. 54%, P = 0.02), and frequency of cavitation on chest CT (12% vs. 68%, P < 0.01). Our findings should raise awareness that neither a positive culture nor the hallmark symptoms are invariably associated with early TB disease. PMID:26448182

  2. Cytological value of sputum in workers daily exposed to air pollution.

    PubMed

    Alderisio, Mauro; Cenci, Maria; Mudu, Pierpaolo; Vecchione, Aldo; Giovagnoli, Maria Rosaria

    2006-01-01

    In this study, quantitative modifications of dust cells, siderocytes, Curschmann's spirals and asbestos bodies and qualitative modifications (cellular changes and inflammatory infiltrate) in the sputum of 164 traffic police officers and 218 railway workers, occupationally exposed to environmental pollution, and the sputum of 119 inhabitants of a rural area, were evaluated. The results were correlated with time of exposure and smoking habits. Seventy-three (45%) traffic police officers (TPO), 76 (35%) railway workers (RW) and 29 (24%) of the rural population (RP) were smokers. The sputum, collected over a 3-day period, was smeared on glass slides and stained according to the Papanicolaou, Perl and yellow eosin methods. The results of the qualitative cytological diagnosis revealed a statistically significant difference between the TPO, RW and the RP (p < 0.001). The results of the qualitative and quantitative cytological examinations were not significantly correlated to time of occupational exposure, which was considered to be a continuous variable. The qualitative cytological examination of sputa was not statistically significant for the smoking habits of the TPO and the RP, but was significant for the RW (p < 0.0067). In the TPO, the number of dust cells was higher in smokers, and the relative risk (RR) was 3.95. In the RW, the RR was 2.84. The results of our study revealed that for the RW, the qualitative-quantitative cytological alterations in sputum were due much more to smoking habits than to occupational exposure, while the presence of asbestos bodies correlated with work activity. The qualitative-quantitative cytological examinations of the TPO differed significantly from that of the other two populations.

  3. Destruction-free procedure for the isolation of bacteria from sputum samples for Raman spectroscopic analysis.

    PubMed

    Kloß, Sandra; Lorenz, Björn; Dees, Stefan; Labugger, Ines; Rösch, Petra; Popp, Jürgen

    2015-11-01

    Lower respiratory tract infections are the fourth leading cause of death worldwide. Here, a timely identification of the causing pathogens is crucial to the success of the treatment. Raman spectroscopy allows for quick identification of bacterial cells without the need for time-consuming cultivation steps, which is the current gold standard to detect pathogens. However, before Raman spectroscopy can be used to identify pathogens, they have to be isolated from the sample matrix, i.e., sputum in case of lower respiratory tract infections. In this study, we report an isolation protocol for single bacterial cells from sputum samples for Raman spectroscopic identification. Prior to the isolation, a liquefaction step using the proteolytic enzyme mixture Pronase E is required in order to deal with the high viscosity of sputum. The extraction of the bacteria was subsequently performed via different filtration and centrifugation steps, whereby isolation ratios between 46 and 57 % were achieved for sputa spiked with 6·10(7) to 6·10(4) CFU/mL of Staphylococcus aureus. The compatibility of such a liquefaction and isolation procedure towards a Raman spectroscopic classification was shown for five different model species, namely S. aureus, Staphylococcus epidermidis, Streptococcus pneumoniae, Klebsiella pneumoniae, and Pseudomonas aeruginosa. A classification of single-cell Raman spectra of these five species with an accuracy of 98.5 % could be achieved on the basis of a principal component analysis (PCA) followed by a linear discriminant analysis (LDA). These classification results could be validated with an independent test dataset, where 97.4 % of all spectra were identified correctly. Graphical Abstract Development of an isolation protocol of bacterial cells out of sputum samples followed by Raman spectroscopic measurement and species identification using chemometrical models. PMID:26041453

  4. Human Neutrophils Secrete Bioactive Paucimannosidic Proteins from Azurophilic Granules into Pathogen-Infected Sputum*

    PubMed Central

    Thaysen-Andersen, Morten; Venkatakrishnan, Vignesh; Loke, Ian; Laurini, Christine; Diestel, Simone; Parker, Benjamin L.; Packer, Nicolle H.

    2015-01-01

    Unlike plants and invertebrates, mammals reportedly lack proteins displaying asparagine (N)-linked paucimannosylation (mannose1–3fucose0–1N-acetylglucosamine2Asn). Enabled by technology advancements in system-wide biomolecular characterization, we document that protein paucimannosylation is a significant host-derived molecular signature of neutrophil-rich sputum from pathogen-infected human lungs and is negligible in pathogen-free sputum. Five types of paucimannosidic N-glycans were carried by compartment-specific and inflammation-associated proteins of the azurophilic granules of human neutrophils including myeloperoxidase (MPO), azurocidin, and neutrophil elastase. The timely expressed human azurophilic granule-resident β-hexosaminidase A displayed the capacity to generate paucimannosidic N-glycans by trimming hybrid/complex type N-glycan intermediates with relative broad substrate specificity. Paucimannosidic N-glycoepitopes showed significant co-localization with β-hexosaminidase A and the azurophilic marker MPO in human neutrophils using immunocytochemistry. Furthermore, promyelocyte stage-specific expression of genes coding for paucimannosidic proteins and biosynthetic enzymes indicated a novel spatio-temporal biosynthetic route in early neutrophil maturation. The absence of bacterial exoglycosidase activities and paucimannosidic N-glycans excluded exogenous origins of paucimannosylation. Paucimannosidic proteins from isolated and sputum neutrophils were preferentially secreted upon inoculation with virulent Pseudomonas aeruginosa. Finally, paucimannosidic proteins displayed affinities to mannose-binding lectin, suggesting immune-related functions of paucimannosylation in activated human neutrophils. In conclusion, we are the first to document that human neutrophils produce, store and, upon activation, selectively secrete bioactive paucimannosidic proteins into sputum of lungs undergoing pathogen-based inflammation. PMID:25645918

  5. Human neutrophils secrete bioactive paucimannosidic proteins from azurophilic granules into pathogen-infected sputum.

    PubMed

    Thaysen-Andersen, Morten; Venkatakrishnan, Vignesh; Loke, Ian; Laurini, Christine; Diestel, Simone; Parker, Benjamin L; Packer, Nicolle H

    2015-04-01

    Unlike plants and invertebrates, mammals reportedly lack proteins displaying asparagine (N)-linked paucimannosylation (mannose(1-3)fucose(0-1)N-acetylglucosamine(2)Asn). Enabled by technology advancements in system-wide biomolecular characterization, we document that protein paucimannosylation is a significant host-derived molecular signature of neutrophil-rich sputum from pathogen-infected human lungs and is negligible in pathogen-free sputum. Five types of paucimannosidic N-glycans were carried by compartment-specific and inflammation-associated proteins of the azurophilic granules of human neutrophils including myeloperoxidase (MPO), azurocidin, and neutrophil elastase. The timely expressed human azurophilic granule-resident β-hexosaminidase A displayed the capacity to generate paucimannosidic N-glycans by trimming hybrid/complex type N-glycan intermediates with relative broad substrate specificity. Paucimannosidic N-glycoepitopes showed significant co-localization with β-hexosaminidase A and the azurophilic marker MPO in human neutrophils using immunocytochemistry. Furthermore, promyelocyte stage-specific expression of genes coding for paucimannosidic proteins and biosynthetic enzymes indicated a novel spatio-temporal biosynthetic route in early neutrophil maturation. The absence of bacterial exoglycosidase activities and paucimannosidic N-glycans excluded exogenous origins of paucimannosylation. Paucimannosidic proteins from isolated and sputum neutrophils were preferentially secreted upon inoculation with virulent Pseudomonas aeruginosa. Finally, paucimannosidic proteins displayed affinities to mannose-binding lectin, suggesting immune-related functions of paucimannosylation in activated human neutrophils. In conclusion, we are the first to document that human neutrophils produce, store and, upon activation, selectively secrete bioactive paucimannosidic proteins into sputum of lungs undergoing pathogen-based inflammation. PMID:25645918

  6. Sputum Mycobacterium tuberculosis mRNA as a marker of bacteriologic clearance in response to antituberculosis therapy.

    PubMed

    Li, L; Mahan, C S; Palaci, M; Horter, L; Loeffelholz, L; Johnson, J L; Dietze, R; Debanne, S M; Joloba, M L; Okwera, A; Boom, W H; Eisenach, K D

    2010-01-01

    mRNA is a marker of cell viability. Quantifying Mycobacterium tuberculosis mRNA in sputum is a promising tool for monitoring response to antituberculosis therapy and evaluating the efficacy of individual drugs. mRNA levels were measured in sputum specimens from patients with tuberculosis (TB) receiving monotherapy in an early bactericidal activity study of fluoroquinolones and in those receiving a standard rifampin-based regimen in an interleukin-2 (IL-2) trial. In the early bactericidal activity study, sputum for quantitative culture and mRNA analysis was collected for 2 days before and daily during 7 days of study drug administration. In the IL-2 trial, sputum was collected for quantitative culture, Bactec 460 liquid culture, and mRNA analysis throughout the intensive treatment phase. RNA was isolated from digested sputum and tested in quantitative reverse transcription-PCR assays for several gene targets. mRNA for the glyoxylate cycle enzyme isocitrate lyase declined at similar rates in patients receiving isoniazid, gatifloxicin, levofloxacin, and moxifloxacin monotherapy. Isocitrate lyase mRNA correlated highly with CFU in sputum prior to therapy and during 7 days of monotherapy in all treatment arms. Isocitrate lyase mRNA was detectable in sputum of culture-positive TB patients receiving a rifampin-based regimen for 1 month. At 2 months, sputum for isocitrate mRNA correlated more closely with growth in liquid culture than did growth on solid culture medium. Data suggest that isocitrate lyase mRNA is a reliable marker of M. tuberculosis viability.

  7. Educational intervention for collecting sputum for tuberculosis: a quasi-experimental study 1

    PubMed Central

    Sicsú, Amélia Nunes; Salem, Julia Ignez; Fujimoto, Luciana Botinelly Mendonça; Gonzales, Roxana Isabel Cardozo; Cardoso, Maria do Socorro de Lucena; Palha, Pedro Fredemir

    2016-01-01

    Abstract Objective: to evaluate the quality of the sputum sample before and after the Nursing guidance to patients. Methods: this is a quasi-experimental research design, single group type, before and after, non-randomized study. The study enrolled patients with suspected pulmonary tuberculosis, respiratory symptomatic patients for over three weeks, aged over 18 years, of both genders and without tuberculosis history in the last two years. The educational intervention consisted of individualized guidance on the collection of sputum sample, which was based on the guidelines of the Ministry of Health of Brazil and on the explanatory folder delivery. Results: in this study participated 138 patients with suspected pulmonary tuberculosis. The results showed significant increase of the samples with purulent particles, volume greater than 5 mL and increased rate of patients diagnosed with tuberculosis, after the educational intervention. Conclusion: it was shown that after the educational intervention, it was observed sputum samples with better quality, with satisfactory aspect and volume for the effectiveness of the bacilloscopic examination. PMID:27276015

  8. Sputum proteomics identifies elevated PIGR levels in smokers and mild-to-moderate COPD.

    PubMed

    Ohlmeier, Steffen; Mazur, Witold; Linja-Aho, Anna; Louhelainen, Noora; Rönty, Mikko; Toljamo, Tuula; Bergmann, Ulrich; Kinnula, Vuokko L

    2012-02-01

    Chronic obstructive pulmonary disease (COPD) is one of the leading causes of morbidity and mortality around the world. However, the exact mechanisms leading to COPD and its progression are still poorly understood. In this study, induced sputum was analyzed by cysteine-specific two-dimensional difference gel electrophoresis (2D-DIGE) coupled with mass spectrometry to identify proteins involved in COPD pathogenesis. The comparison of nonsmokers, smokers, and smokers with moderate COPD revealed 15 changed proteins with the majority, including polymeric immunoglobulin receptor (PIGR), being elevated in smokers and subjects with COPD. PIGR, which is involved in specific immune defense and inflammation, was further studied in sputum, lung tissue, and plasma by Western blot, immunohistochemistry/image analysis, and/or ELISA. Sputum PIGR was characterized as glycosylated secretory component (SC). Lung PIGR was significantly elevated in the bronchial and alveolar epithelium of smokers and further increased in the alveolar area in mild to moderate COPD. Plasma PIGR was elevated in smokers and smokers with COPD compared to nonsmokers with significant correlation to obstruction. In conclusion, new proteins in smoking-related chronic inflammation and COPD could be identified, with SC/PIGR being one of the most prominent not only in the lung but also in circulating blood.

  9. Phenotypically Adapted Mycobacterium tuberculosis Populations from Sputum Are Tolerant to First-Line Drugs

    PubMed Central

    Turapov, Obolbek; O'Connor, Benjamin D.; Sarybaeva, Asel A.; Williams, Caroline; Patel, Hemu; Kadyrov, Abdullaat S.; Sarybaev, Akpay S.; Woltmann, Gerrit; Barer, Michael R.

    2016-01-01

    Tuberculous sputum contains multiple Mycobacterium tuberculosis populations with different requirements for isolation in vitro. These include cells that form colonies on solid media (plateable M. tuberculosis), cells requiring standard liquid medium for growth (nonplateable M. tuberculosis), and cells requiring supplementation of liquid medium with culture supernatant (SN) for growth (SN-dependent M. tuberculosis). Here, we describe protocols for the cryopreservation and direct assessment of antimicrobial tolerance of these M. tuberculosis populations within sputum. Our results show that first-line drugs achieved only modest bactericidal effects on all three populations over 7 days (1 to 2.5 log10 reductions), and SN-dependent M. tuberculosis was more tolerant to streptomycin and isoniazid than the plateable and nonplateable M. tuberculosis strains. Susceptibility of plateable M. tuberculosis to bactericidal drugs was significantly increased after passage in vitro; thus, tolerance observed in the sputum samples from the population groups was likely associated with mycobacterial adaptation to the host environment at some time prior to expectoration. Our findings support the use of a simple ex vivo system for testing drug efficacies against mycobacteria that have phenotypically adapted during tuberculosis infection. PMID:26883695

  10. Ferruginous body counts in sputum as an index of past exposure to mineral fibres.

    PubMed

    McDonald, J C; Sébastien, P; Case, B; McDonald, A D; Dufresne, A

    1992-06-01

    Previous work by our group among vermiculite miners exposed to fibrous tremolite demonstrated that asbestos body counts in sputum closely reflected intensity and duration of past exposure. In the present project sputum samples from nearly 600 volunteers from 11 cohorts of workers exposed to asbestos and other mineral fibres were found to contain much lower numbers of asbestos bodies than had been observed in vermiculite workers and these counts did not reliably reflect past levels of exposure. No evidence was found that occupational exposure to man-made mineral fibres led to any ferruginous body formation. Asbestos body counts appeared to differentiate between mesothelioma cases and controls and between levels of radiological asbestosis, but in both comparisons, based on small numbers, the power of discrimination was low. Substantial variation was found both in asbestos body and in macrophage counts in daily sampling of vermiculite workers but it was not sufficient to invalidate comparison between groups for epidemiological study. In individual subjects, however, little reliance can be put on results from a single sputum sample, particularly if negative.

  11. Impact of storage conditions on metabolite profiles of sputum samples from persons with cystic fibrosis

    PubMed Central

    Zhao, Jiangchao; Evans, Charles R.; Carmody, Lisa A.; LiPuma, John J.

    2016-01-01

    Background Although recent studies have begun to elucidate how airway microbial community structure relates to lung disease in cystic fibrosis (CF), microbial community activity and the host’s response to changes in this activity are poorly understood. Metabolomic profiling provides a means to investigate microbial and human cell activity within diseased airways. However, variables in sample storage and shipping likely affect downstream analyses and standards for sample handling are lacking. Methods We assessed the impact of sample storage conditions on liquid chromatography mass spectrometry analysis of CF sputum samples. Results Significant changes in global metabolomic profiles occurred in samples stored at room temperature or at 4°C for longer than one day. Untargeted metabolomic profiles were stable in sputum samples stored at −20°C or −80°C for at least 28 days. Quorum sensing molecules and phenazines, both considered important to the in vivo activity of Pseudomonas during airway infection, were detected after sample storage at room temperature for five days. Conclusions Sputum samples can be stored at −20°C or −80°C for weeks with minimal effect on global metabolomic profiles. This observation provides guidance in designing metabolomic studies that have the potential to deepen our understanding of how airway microbial communities impact lung disease progression in CF. PMID:25725986

  12. Analysis of the association between host genetics, smoking, and sputum microbiota in healthy humans

    PubMed Central

    Lim, Mi Young; Yoon, Hyo Shin; Rho, Mina; Sung, Joohon; Song, Yun-Mi; Lee, Kayoung; Ko, GwangPyo

    2016-01-01

    Recent studies showing clear differences in the airway microbiota between healthy and diseased individuals shed light on the importance of the airway microbiota in health. Here, we report the associations of host genetics and lifestyles such as smoking, alcohol consumption, and physical activity with the composition of the sputum microbiota using 16S rRNA gene sequence data generated from 257 sputum samples of Korean twin-family cohort. By estimating the heritability of each microbial taxon, we found that several taxa, including Providencia and Bacteroides, were significantly influenced by host genetic factors. Smoking had the strongest effect on the overall microbial community structure among the tested lifestyle factors. The abundances of Veillonella and Megasphaera were higher in current-smokers, and increased with the pack-year value and the Fagerstrom Test of Nicotine Dependence (FTND) score. In contrast, Haemophilus decreased with the pack-year of smoking and the FTND score. Co-occurrence network analysis showed that the taxa were clustered according to the direction of associations with smoking, and that the taxa influenced by host genetics were found together. These results demonstrate that the relationships among sputum microbial taxa are closely associated with not only smoking but also host genetics. PMID:27030383

  13. Analysis of the association between host genetics, smoking, and sputum microbiota in healthy humans.

    PubMed

    Lim, Mi Young; Yoon, Hyo Shin; Rho, Mina; Sung, Joohon; Song, Yun-Mi; Lee, Kayoung; Ko, GwangPyo

    2016-03-31

    Recent studies showing clear differences in the airway microbiota between healthy and diseased individuals shed light on the importance of the airway microbiota in health. Here, we report the associations of host genetics and lifestyles such as smoking, alcohol consumption, and physical activity with the composition of the sputum microbiota using 16S rRNA gene sequence data generated from 257 sputum samples of Korean twin-family cohort. By estimating the heritability of each microbial taxon, we found that several taxa, including Providencia and Bacteroides, were significantly influenced by host genetic factors. Smoking had the strongest effect on the overall microbial community structure among the tested lifestyle factors. The abundances of Veillonella and Megasphaera were higher in current-smokers, and increased with the pack-year value and the Fagerstrom Test of Nicotine Dependence (FTND) score. In contrast, Haemophilus decreased with the pack-year of smoking and the FTND score. Co-occurrence network analysis showed that the taxa were clustered according to the direction of associations with smoking, and that the taxa influenced by host genetics were found together. These results demonstrate that the relationships among sputum microbial taxa are closely associated with not only smoking but also host genetics.

  14. Analysis of the association between host genetics, smoking, and sputum microbiota in healthy humans.

    PubMed

    Lim, Mi Young; Yoon, Hyo Shin; Rho, Mina; Sung, Joohon; Song, Yun-Mi; Lee, Kayoung; Ko, GwangPyo

    2016-01-01

    Recent studies showing clear differences in the airway microbiota between healthy and diseased individuals shed light on the importance of the airway microbiota in health. Here, we report the associations of host genetics and lifestyles such as smoking, alcohol consumption, and physical activity with the composition of the sputum microbiota using 16S rRNA gene sequence data generated from 257 sputum samples of Korean twin-family cohort. By estimating the heritability of each microbial taxon, we found that several taxa, including Providencia and Bacteroides, were significantly influenced by host genetic factors. Smoking had the strongest effect on the overall microbial community structure among the tested lifestyle factors. The abundances of Veillonella and Megasphaera were higher in current-smokers, and increased with the pack-year value and the Fagerstrom Test of Nicotine Dependence (FTND) score. In contrast, Haemophilus decreased with the pack-year of smoking and the FTND score. Co-occurrence network analysis showed that the taxa were clustered according to the direction of associations with smoking, and that the taxa influenced by host genetics were found together. These results demonstrate that the relationships among sputum microbial taxa are closely associated with not only smoking but also host genetics. PMID:27030383

  15. Atmospheric remote sensing to detect effects of temperature inversions on sputum cell counts in airway diseases.

    PubMed

    Wallace, Julie; Nair, Parameswaran; Kanaroglou, Pavlos

    2010-08-01

    Temperature inversions result in the accumulation of air pollution, often to levels exceeding air quality criteria. The respiratory response may be detectable in sputum cell counts. This study investigates the effect of boundary layer temperature inversions on sputum cell counts. Total and differential cell counts of neutrophils, eosinophils, macrophages and lymphocytes were quantified in sputum samples of patients attending an outpatient clinic. Temperature inversions were identified using data from the Atmospheric Infrared Sounder, an atmospheric sensor on the Aqua spacecraft which was launched in 2002 by the National Aeronautics and Space Administration. On inversion days, a statistically significant increase in the percent of cells that were neutrophils was observed in stable patients. There was also a statistically significant increase in the percent of cells that were macrophages, in exacerbated patients. Multivariate linear regression models were used to assess the relationship between temperature inversions and cell counts, controlling patients' age, smoking status, medications and meteorological variables of temperature and humidity. The analyses indicate that, in the stable and exacerbated groups, percent neutrophils and macrophages increased by 12.6% and 2.5%, respectively, on inversion days. These results suggest that temperature inversions need consideration as an exacerbating factor in bronchitis and obstructive airway disease. The effects of air pollutants, nitrogen dioxide, carbon monoxide, fine particulate matter and ozone, were investigated. We identified no significant associations with any pollutant. However, we found that monthly averages of total cell counts were strongly correlated with monthly nitrogen dioxide concentrations, an association not previously identified in the literature.

  16. Microbiological profiles of sputum and gastric juice aspirates in Cystic Fibrosis patients

    PubMed Central

    Al-momani, H.; Perry, A.; Stewart, C. J.; Jones, R.; Krishnan, A.; Robertson, A. G.; Bourke, S.; Doe, S.; Cummings, S. P.; Anderson, A.; Forrest, T.; Griffin, S. M.; Brodlie, M.; Pearson, J.; Ward, C.

    2016-01-01

    Gastro-Oesophageal Reflux (GOR) is a key problem in Cystic Fibrosis (CF), but the relationship between lung and gastric microbiomes is not well understood. We hypothesised that CF gastric and lung microbiomes are related. Gastric and sputum cultures were obtained from fifteen CF patients receiving percutaneous endoscopic gastrostomy feeding. Non-CF gastric juice data was obtained through endoscopy from 14 patients without lung disease. Bacterial and fungal isolates were identified by culture. Molecular bacterial profiling used next generation sequencing (NGS) of the 16S rRNA gene. Cultures grew bacteria and/or fungi in all CF gastric juice and sputa and in 9/14 non-CF gastric juices. Pseudomonas aeruginosa(Pa) was present in CF sputum in 11 patients, 4 had identical Pa strains in the stomach. NGS data from non-CF gastric juice samples were significantly more diverse compared to CF samples. NGS showed CF gastric juice had markedly lower abundance of normal gut bacteria; Bacteroides and Faecalibacterium, but increased Pseudomonas compared with non-CF. Multivariate partial least squares discriminant analysis demonstrated similar bacterial profiles of CF sputum and gastric juice samples, which were distinct from non-CF gastric juice. We provide novel evidence suggesting the existence of an aerodigestive microbiome in CF, which may have clinical relevance. PMID:27245316

  17. Nedocromil sodium in obstructive airways disease: effect on symptoms and plasma protein leakage in sputum.

    PubMed

    Schoonbrood, D F; Out, T A; Hart, A A; Habets, F J; Roos, C M; Jansen, H M

    1997-07-01

    In patients with asthma or chronic obstructive pulmonary disease, there is chronic airway inflammation with increased leakage of plasma proteins into the airway lumen, which can be reduced by inhaled glucocorticosteroids. Nedocromil sodium is an anti-inflammatory drug, and we questioned whether it also affects the leakage of plasma proteins. In a double-blind placebo-controlled study we investigated the effect of 12 weeks of treatment with nedocromil on forced expiratory volume in one second (FEV1), provocative concentration of histamine causing a 20% fall in FEV1 (PC20), peak flow, symptom scores, and plasma protein leakage in sputum, in 31 patients with obstructive airways disease and sputum production (mean (range) FEV1 61% of predicted (42-87%); geometric mean (range) PC20 0.39 (0.04-2.9) mg x mL(-1)). As a measure for plasma protein leakage we calculated the relative coefficients of excretion (RCE) of proteins from serum to the soluble phase of sputum. There was a small increase in morning and evening peak flow (p<0.05) and a decrease in night-time bronchodilator use (p<0.02) in favour of nedocromil. The RCE of alpha2-macroglobulin to albumin significantly decreased after treatment with nedocromil (p=0.03). The results show limited clinical efficacy of nedocromil in our study group. They further suggest that the anti-inflammatory properties of nedocromil extend to inhibition of plasma protein leakage into the airways.

  18. Pilot study of a rapid and minimally instrumented sputum sample preparation method for molecular diagnosis of tuberculosis.

    PubMed

    Ferguson, Tanya M; Weigel, Kris M; Lakey Becker, Annie; Ontengco, Delia; Narita, Masahiro; Tolstorukov, Ilya; Doebler, Robert; Cangelosi, Gerard A; Niemz, Angelika

    2016-01-01

    Nucleic acid amplification testing (NAAT) enables rapid and sensitive diagnosis of tuberculosis (TB), which facilitates treatment and mitigates transmission. Nucleic acid extraction from sputum constitutes the greatest technical challenge in TB NAAT for near-patient settings. This report presents preliminary data for a semi-automated sample processing method, wherein sputum is disinfected and liquefied, followed by PureLyse(®) mechanical lysis and solid-phase nucleic acid extraction in a miniaturized, battery-operated bead blender. Sputum liquefaction and disinfection enabled a >10(4) fold reduction in viable load of cultured Mycobacterium tuberculosis (M.tb) spiked into human sputum, which mitigates biohazard concerns. Sample preparation via the PureLyse(®) method and a clinically validated manual method enabled positive PCR-based detection for sputum spiked with 10(4) and 10(5) colony forming units (cfu)/mL M.tb. At 10(3) cfu/mL sputum, four of six and two of six samples amplified using the comparator and PureLyse(®) method, respectively. For clinical specimens from TB cases and controls, the two methods provided 100% concordant results for samples with 1 mL input volume (N = 41). The semi-automated PureLyse(®) method therefore performed similarly to a validated manual comparator method, but is faster, minimally instrumented, and can be integrated into TB molecular diagnostic platforms designed for near-patient low-resource settings. PMID:26785769

  19. Radiation inactivation of Paenibacillus larvae and sterilization of American Foul Brood (AFB) infected hives using Co-60 gamma rays.

    PubMed

    De Guzman, Zenaida M; Cervancia, Cleofas R; Dimasuay, Kris Genelyn B; Tolentino, Mitos M; Abrera, Gina B; Cobar, Ma Lucia C; Fajardo, Alejandro C; Sabino, Noel G; Manila-Fajardo, Analinda C; Feliciano, Chitho P

    2011-10-01

    The effectiveness of gamma radiation in inactivating the Philippine isolate of Paenibacillus larvae was investigated. Spores of P. larvae were irradiated at incremental doses (0.1, 0.2, 0.4, 0.8 and 1.6 kGy) of gamma radiation emitted by a ⁶⁰Co source. Surviving spores were counted and used to estimate the decimal reduction (D₁₀) value. A dose of 0.2 kGy was sufficient to inactivate 90% of the total recoverable spores from an initial count of 10⁵- 9 × 10³ spores per glass plate. The sterilizing effect of high doses of gamma radiation on the spores of P. larvae in infected hives was determined. In this study, a minimum dose (D(min)) of 15 kGy was tested. Beehives with sub-clinical infections of AFB were irradiated and examined for sterility. All the materials were found to be free of P. larvae indicating its susceptibility to γ-rays. After irradiation, there were no visible changes in the physical appearance of the hives' body, wax and frames. Thus, a dose of 15 kGy is effective enough for sterilization of AFB-infected materials.

  20. Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element

    PubMed Central

    Ertekin, Özlem; Öztürk, Selma; Öztürk, Zafer Ziya

    2016-01-01

    This study introduces the use of an IgA isotype aflatoxin (AF) specific monoclonal antibody for the development of a highly sensitive Quartz Crystal Microbalance (QCM) immunobiosensor for the detection of AF in inhibitory immunoassay format. The higher molecular weight of IgA antibodies proved an advantage over commonly used IgG antibodies in label free immunobiosensor measurements. IgA and IgG antibodies with similar affinity for AF were used in the comparative studies. Sensor surface was prepared by covalent immobilization of AFB1, using self assembled monolayer (SAM) formed on gold coated Quartz Crystal, with 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxy succinimide (EDC/NHS) method using a diamine linker. Nonspecific binding to the surface was decreased by minimizing the duration of EDC/NHS activation. Sensor surface was chemically blocked after AF immobilization without any need for protein blocking. This protein free sensor chip endured harsh solutions with strong ionic detergent at high pH, which is required for the regeneration of the high affinity antibody-antigen interaction. According to the obtained results, the detection range with IgA antibodies was higher than IgG antibodies in QCM immunosensor developed for AFB1. PMID:27529243

  1. Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element.

    PubMed

    Ertekin, Özlem; Öztürk, Selma; Öztürk, Zafer Ziya

    2016-01-01

    This study introduces the use of an IgA isotype aflatoxin (AF) specific monoclonal antibody for the development of a highly sensitive Quartz Crystal Microbalance (QCM) immunobiosensor for the detection of AF in inhibitory immunoassay format. The higher molecular weight of IgA antibodies proved an advantage over commonly used IgG antibodies in label free immunobiosensor measurements. IgA and IgG antibodies with similar affinity for AF were used in the comparative studies. Sensor surface was prepared by covalent immobilization of AFB1, using self assembled monolayer (SAM) formed on gold coated Quartz Crystal, with 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxy succinimide (EDC/NHS) method using a diamine linker. Nonspecific binding to the surface was decreased by minimizing the duration of EDC/NHS activation. Sensor surface was chemically blocked after AF immobilization without any need for protein blocking. This protein free sensor chip endured harsh solutions with strong ionic detergent at high pH, which is required for the regeneration of the high affinity antibody-antigen interaction. According to the obtained results, the detection range with IgA antibodies was higher than IgG antibodies in QCM immunosensor developed for AFB1. PMID:27529243

  2. Fluoro-edenitic fibres in the sputum of subjects from Biancavilla (Sicily): a pilot study

    PubMed Central

    Putzu, Maria Grazia; Bruno, Caterina; Zona, Amerigo; Massiccio, Marilena; Pasetto, Roberto; Piolatto, Pier Giorgio; Comba, Pietro

    2006-01-01

    Background An excess of mortality for malignant neoplasms of the pleura in Biancavilla, promoted an investigation for pleural mesothelioma, disclosing 17 cases. As the absence of known sources of asbestos exposure, a local stone quarry, located near the inhabited area, used for the extraction of building materials, was investigated. Amphibolic fibres were found in the quarry and identified as fluoro-edenite "new end-member of the edenite / fluoro-edenite series" and recognized as the fluoro-edenite holotype by International Mineralogical Association – Commission on New Minerals and Mineral Names. A pilot study was performed to verify the feasibility of using spontaneous sputum as an exposure indicator for these fibres, in a context in which the use of aerosol-induced sputum technique would not be easily accepted. Methods Hypothesizing a behaviour of the new fibre analogous to that of asbestos, the determination of the free fibres and the ferruginous bodies in spontaneous sputum was carried out. Phase Contrast Optical Microscope and an Environmental Scanning Electron Microscope fitted with X-ray energy dispersive analysis system (micro-analysis) were used to examine the samples. The criteria for inclusion in the study were: 1) subjects hospitalized for exacerbation of chronic obstructive pulmonary disease symptoms, 2) age ≥ 45 years, 3) residence in Biancavilla for at least 30 years. Results The preliminary findings are related to 12 subjects (7 females and 5 males). Uncoated fibres (with length > 5 μm, diameter < 3 μm, aspect ratio 3.1) and ferruginous bodies were searched. Six out of twelve subjects (4 females, 2 males) had at least one of the three samples positive for the presence of fluoro-edenite, confirmed by micro-analysis. The fibre concentration found in the sputum ranged from 0.04 to 10 fibres/g; the length from 20 to 40 μm, the diameter was < 0.5 μm. No ferruginous bodies were found in any of the samples. The four women with a positive sample

  3. Sputum Inflammatory Profile Before and After Specific Inhalation Challenge in Individuals with Suspected Occupational Asthma

    PubMed Central

    Sánchez-Vidaurre, Sara; Cruz, María-Jesús; Gómez-Ollés, Susana; Morell, Ferran; Muñoz, Xavier

    2013-01-01

    Background The aim of this study was to establish the sputum inflammatory profile and changes in levels of leukotriene B4 (LTB4) and a panel of Th1/Th2 cytokines in subjects with suspected occupational asthma (OA) following specific inhalation challenge (SIC) to high-molecular-weight (HMW) and low-molecular-weight (LMW) agents. Material and Methods Fifty-one consecutive subjects undergoing SIC for suspected OA were enrolled. Sputum induction was performed the day before and 24 h after exposure to the offending agent. Total and differential cell counts were assessed. LTB4 and a 10 Th1/Th2 cytokines were measured in sputum supernatant. Results Thirty-four patients tested positive to SIC and were diagnosed with OA (in 10 due to HMW agents and in 24 to LMW agents). SIC was negative in 17 subjects. As compared to baseline an increase was found in the percentage of sputum eosinophils and neutrophils, and in IL-10 concentration after SIC (p = 0.0078, p = 0.0195, and p = 0.046, respectively), and a decrease was seen in LTB4 level (p = 0.0078) in patients with OA due to HMW agents. An increase in the percentage of sputum neutrophils after SIC (p = 0.0040) was observed in subjects without OA exposed to LMW agents. IL-8 levels after SIC were higher in patients without OA compared with patients with OA (p = 0.0146). Conclusion When conducting airway inflammation studies in OA, patients should be divided according to the causal agent (HMW or LMW). In OA patients exposed to HMW agents, an increase in the number of neutrophils can be found in parallel to the increase of eosinophils, although this does not contradict an IgE-mediated mechanism. Exposure to LMW agents can result in increased neutrophilic inflammation in patients with airway diseases unrelated to OA. There is variability in the responses observed in patients with OA exposed to LMW agents. PMID:24236015

  4. Compliance testing of Eielson AFB (Air Force Base) central heating and power plant, coal-fired Boiler Number 4, Eielson AFB, Arkansas. Final report, 7-15 June 1989

    SciTech Connect

    Garrison, J.A.

    1989-07-01

    At the request of HQ 343 CSG/DEEV and HQ AAC/SGPB, source compliance testing (particulate and visible emissions) of Boiler No 4 in the Eielson AFB Central Heating and Power Plant was conducted on 7-15 June 1989. Testing was performed to determine compliance with regards to the renewal of Alaska Department of Environmental Conservation Air Quality Control Permit to Operate 8331-AA001. Boiler No. 4 was tested on 12 and 14 Jun 89. On 12 June results indicated that visible-emissions standards were met; however, particulate-emissions standards were not met. The boiler was retested on 14 June and all emissions standards were met. All emission requirements for permit renewal have now been met.

  5. Early and Efficient Detection of Mycobacterium tuberculosis in Sputum by Microscopic Observation of Broth Cultures

    PubMed Central

    Kidenya, Benson R.; Kabangila, Rodrick; Peck, Robert N.; Mshana, Stephen E.; Webster, Lauren E.; Koenig, Serena P.; Johnson, Warren D.; Fitzgerald, Daniel W.

    2013-01-01

    Early, efficient and inexpensive methods for the detection of pulmonary tuberculosis are urgently needed for effective patient management as well as to interrupt transmission. These methods to detect M. tuberculosis in a timely and affordable way are not yet widely available in resource-limited settings. In a developing-country setting, we prospectively evaluated two methods for culturing and detecting M. tuberculosis in sputum. Sputum samples were cultured in liquid assay (micro broth culture) in microplate wells and growth was detected by microscopic observation, or in Löwenstein–Jensen (LJ) solid media where growth was detected by visual inspection for colonies. Sputum samples were collected from 321 tuberculosis (TB) suspects attending Bugando Medical Centre, in Mwanza, Tanzania, and were cultured in parallel. Pulmonary tuberculosis cases were diagnosed using the American Thoracic Society diagnostic standards. There were a total of 200 (62.3%) pulmonary tuberculosis cases. Liquid assay with microscopic detection detected a significantly higher proportion of cases than LJ solid culture: 89.0% (95% confidence interval [CI], 84.7% to 93.3%) versus 77.0% (95% CI, 71.2% to 82.8%) (p = 0.0007). The median turn around time to diagnose tuberculosis was significantly shorter for micro broth culture than for the LJ solid culture, 9 days (interquartile range [IQR] 7–13), versus 21 days (IQR 14–28) (p<0.0001). The cost for micro broth culture (labor inclusive) in our study was US $4.56 per sample, versus US $11.35 per sample for the LJ solid culture. The liquid assay (micro broth culture) is an early, feasible, and inexpensive method for detection of pulmonary tuberculosis in resource limited settings. PMID:23469014

  6. Sputum containing zinc enhances carbapenem resistance, biofilm formation and virulence of Pseudomonas aeruginosa.

    PubMed

    Marguerettaz, Mélanie; Dieppois, Guennaëlle; Que, Yok Ai; Ducret, Véréna; Zuchuat, Sandrine; Perron, Karl

    2014-12-01

    Pseudomonas aeruginosa chronic lung infections are the leading cause of mortality in cystic fibrosis patients, a serious problem which is notably due to the numerous P. aeruginosa virulence factors, to its ability to form biofilms and to resist the effects of most antibiotics. Production of virulence factors and biofilm formation by P. aeruginosa is highly coordinated through complex regulatory systems. We recently found that CzcRS, the zinc and cadmium-specific two-component system is not only involved in metal resistance, but also in virulence and carbapenem antibiotic resistance in P. aeruginosa. Interestingly, zinc has been shown to be enriched in the lung secretions of cystic fibrosis patients. In this study, we investigated whether zinc might favor P. aeruginosa pathogenicity using an artificial sputum medium to mimic the cystic fibrosis lung environment. Our results show that zinc supplementation triggers a dual P. aeruginosa response: (i) it exacerbates pathogenicity by a CzcRS two-component system-dependent mechanism and (ii) it stimulates biofilm formation by a CzcRS-independent mechanism. Furthermore, P. aeruginosa cells embedded in these biofilms exhibited increased resistance to carbapenems. We identified a novel Zn-sensitive regulatory circuit controlling the expression of the OprD porin and modifying the carbapenem resistance profile. Altogether our data demonstrated that zinc levels in the sputum of cystic fibrosis patients might aggravate P. aeruginosa infection. Targeting zinc levels in sputum would be a valuable strategy to curb the increasing burden of P. aeruginosa infections in cystic fibrosis patients. PMID:25448466

  7. Noninvasive Analysis of the Sputum Transcriptome Discriminates Clinical Phenotypes of Asthma

    PubMed Central

    Yan, Xiting; Chu, Jen-Hwa; Gomez, Jose; Koenigs, Maria; Holm, Carole; He, Xiaoxuan; Perez, Mario F.; Zhao, Hongyu; Mane, Shrikant; Martinez, Fernando D.; Ober, Carole; Nicolae, Dan L.; Barnes, Kathleen C.; London, Stephanie J.; Gilliland, Frank; Weiss, Scott T.; Raby, Benjamin A.; Cohn, Lauren

    2015-01-01

    Rationale: The airway transcriptome includes genes that contribute to the pathophysiologic heterogeneity seen in individuals with asthma. Objectives: We analyzed sputum gene expression for transcriptomic endotypes of asthma (TEA), gene signatures that discriminate phenotypes of disease. Methods: Gene expression in the sputum and blood of patients with asthma was measured using Affymetrix microarrays. Unsupervised clustering analysis based on pathways from the Kyoto Encyclopedia of Genes and Genomes was used to identify TEA clusters. Logistic regression analysis of matched blood samples defined an expression profile in the circulation to determine the TEA cluster assignment in a cohort of children with asthma to replicate clinical phenotypes. Measurements and Main Results: Three TEA clusters were identified. TEA cluster 1 had the most subjects with a history of intubation (P = 0.05), a lower prebronchodilator FEV1 (P = 0.006), a higher bronchodilator response (P = 0.03), and higher exhaled nitric oxide levels (P = 0.04) compared with the other TEA clusters. TEA cluster 2, the smallest cluster, had the most subjects that were hospitalized for asthma (P = 0.04). TEA cluster 3, the largest cluster, had normal lung function, low exhaled nitric oxide levels, and lower inhaled steroid requirements. Evaluation of TEA clusters in children confirmed that TEA clusters 1 and 2 are associated with a history of intubation (P = 5.58 × 10−6) and hospitalization (P = 0.01), respectively. Conclusions: There are common patterns of gene expression in the sputum and blood of children and adults that are associated with near-fatal, severe, and milder asthma. PMID:25763605

  8. Diagnostic value of sputum adenosine deaminase (ADA) level in pulmonary tuberculosis

    PubMed Central

    Binesh, Fariba; Jalali, Hadi; Zare, Mohammad Reza; Behravan, Farhad; Tafti, Arefeh Dehghani; Behnaz, Fatemah; Tabatabaee, Mohammad; Shahcheraghi, Seyed Hossein

    2016-01-01

    Introduction Tuberculosis is still a considerable health problem in many countries. Rapid diagnosis of this disease is important, and adenosine deaminase (ADA) has been used as a diagnostic test. The aim of this study was to assess the diagnostic value of ADA in the sputum of patients with pulmonary tuberculosis. Methods The current study included 40 patients with pulmonary tuberculosis (culture positive, smear ±) and 42 patients with non tuberculosis pulmonary diseases (culture negative). ADA was measured on all of the samples. Results The median value of ADA in non-tuberculosis patients was 2.94 (4.2) U/L and 4.01 (6.54) U/L in tuberculosis patients, but this difference was not statistically significant (p=0.100). The cut-off point of 3.1 U/L had a sensitivity of 61% and a specificity of 53%, the cut-off point of 2.81 U/L had a sensitivity of 64% and a specificity of 50% and the cut-off point of 2.78 U/L had a sensitivity of 65% and a specificity of 48%. The positive predictive values for cut-off points of 3.1, 2.81 and 2.78 U/L were 55.7%, 57.44% and 69.23%, respectively. The negative predictive values for the abovementioned cut-off points were 56.75%, 57.14% and 55.88%, respectively. Conclusion Our results showed that sputum ADA test is neither specific nor sensitive. Because of its low sensitivity and specificity, determination of sputum ADA for the diagnosis of pulmonary tuberculosis is not recommended. PMID:27482515

  9. Glutathione-S-transferases in lung and sputum specimens, effects of smoking and COPD severity

    PubMed Central

    Harju, Terttu; Mazur, Witold; Merikallio, Heta; Soini, Ylermi; Kinnula, Vuokko L

    2008-01-01

    Background Oxidative stress plays a potential role in the pathogenesis and progression of chronic obstructive pulmonary disease (COPD). Glutathione S-transferases (GSTs) detoxify toxic compounds in tobacco smoke via glutathione-dependent mechanisms. Little is known about the regulation and expression of GSTs in COPD lung and their presence in airway secretions. Methods GST alpha, pi and mu were investigated by immunohistochemistry in 72 lung tissue specimens and by Western analysis in total lung homogenates and induced sputum supernatants from non-smokers, smokers and patients with variable stages of COPD severity. Results GST alpha was expressed mainly in the airway epithelium. The percentage of GST alpha positive epithelial cells was lower in the central airways of patients with very severe (Stage IV) COPD compared to mild/moderate COPD (p = 0.02). GST alpha by Western analysis was higher in the total lung homogenates in mild/moderate COPD compared to cases of very severe disease (p < 0.001). GST pi was present in airway and alveolar epithelium as well as in alveolar macrophages. GST mu was expressed mainly in the epithelium. Both GST alpha and pi were detectable in sputum supernatants especially in patients with COPD. Conclusion This study indicates the presence of GST alpha and pi especially in the epithelium and sputum supernatants in mild/moderate COPD and low expression of GST alpha in the epithelium in cases of very severe COPD. The presence of GSTs in the airway secretions points to their potential protective role both as intracellular and extracellular mediators in human lung. PMID:19077292

  10. Microparticles in sputum of COPD patients: a potential biomarker of the disease?

    PubMed Central

    Lacedonia, Donato; Carpagnano, Giovanna Elisiana; Trotta, Teresa; Palladino, Grazia Pia; Panaro, Maria Antonietta; Zoppo, Liugi Davide; Foschino Barbaro, Maria Pia; Porro, Chiara

    2016-01-01

    Background Microparticles (MPs) are small membrane vesicles of 0.1–1 µm which are released by cells following chemical, physical, and apoptotic stimuli. MPs represent more than a miniature version of the cell. Their composition and function depend not only on cellular origin, but also on stimuli. Chronic obstructive pulmonary disease (COPD) is a lung disease characterized by nearly irreversible lung destruction which results in airway limitation. Purpose We investigated the presence and source of MPs in sputum of COPD patients to evaluate if changes in MP number and origin may reflect the pathophysiological conditions of disease and may serve as potential biomarkers for diagnostic and prognostic use. Methods Induced sputum samples were collected from 18 male subjects and liquefied with Sputasol. MPs obtained were immunolabeled for leukocyte (CD11a), granulocyte (CD66b), monocyte-macrophage (CD11b), platelets and megakaryocytic cells (CD41), endothelial cells (CD31), and red blood cells (CD235ab) and analyzed by cytofluorimetry. Results There was a negative correlation between CD31-MPs and forced expiratory volume in 1 second (R=−53, P<0.05) and CD66b-MP level was correlated with worse performance index of COPD such as the Body mass index airflow Obstruction, Dyspnea, and Exercise capacity (BODE); they were negatively correlated with 6-minute walking test: 0.65 and −0.64, respectively (P<0.05). CD235ab-MPs showed a negative correlation with body mass index (R=−0.86, P<0.05), while there was a positive correlation with dyspnea index (R=0.91, P<0.05). Conclusion The main finding of this study was that MPs were detected in the sputum of patients affected by COPD. The phenotype of some of them was related to the main COPD parameters. These results suggest that MPs could be implicated in the pathogenesis of COPD. PMID:27042041

  11. Reduced sputum expression of interferon-stimulated genes in severe COPD

    PubMed Central

    Hilzendeger, Clarissa; da Silva, Jane; Henket, Monique; Schleich, Florence; Corhay, Jean Louis; Kebadze, Tatiana; Edwards, Michael R; Mallia, Patrick; Johnston, Sebastian L; Louis, Renaud

    2016-01-01

    Background Exacerbations of COPD are frequent and commonly triggered by respiratory tract infections. The purpose of our study was to investigate innate immunity in stable COPD patients. Methods Induced sputum was collected from 51 stable consecutive COPD patients recruited from the COPD Clinic of CHU Liege and 35 healthy subjects. Expression of interferons beta (IFN-β) and lambda1 (IL-29), IFN-stimulated genes (ISGs) MxA, OAS, and viperin were measured in total sputum cells by reverse transcription quantitative polymerase chain reaction (RT-qPCR). The presence of Picornaviruses was assessed by RT-PCR, while potential pathogenic microorganisms (PPM) were identified by sputum bacteriology. Results Expression of IL-29 was found in 16 of 51 COPD patients (31%) and in nine of 35 healthy subjects (26%), while IFN-β was detected in six of 51 COPD patients (12%) and in two of 35 healthy subjects (6%). ISGs were easily detectable in both groups. In the whole group of COPD patients, OAS expression was decreased (P<0.05), while that of viperin was increased (P<0.01) compared to healthy subjects. No difference was found with respect to MxA. COPD patients from group D of Global Initiative for Chronic Obstructive Lung Disease (GOLD) had reduced expression of all three ISGs (P<0.01 for MxA, P<0.05 for OAS, and P<0.01 for viperin) as compared to those of group B patients. Picornaviruses were detected in eight of 51 (16%) COPD patients vs four of 33 (12%) healthy subjects, while PPM were detected in seven of 39 (18%) COPD patients and associated with raised sputum neutrophil counts. IFN-β expression was raised when either picornavirus or PPM were detected (P=0.06), but no difference was seen regarding IL-29 or ISGs. Conclusion ISGs expression was reduced in severe COPD that may favor exacerbation and contribute to disease progress by altering response to infection. PMID:27418822

  12. Inflammatory Pseudotumor of the Liver with Escherichia coli in the Sputum

    PubMed Central

    Narayan, Sreenath; Nayak, Ashwini; King, Chris L.

    2015-01-01

    Inflammatory pseudotumor is a nonmalignant lesion that mimics malignant lesions and has been reported to occur at various sites throughout the body. Though it has been reported as a reaction to infection, the true etiology of the lesion is unknown. In this report, we present the case of a patient with a liver lesion of unknown origin. Through a series of imaging studies, we were able to observe the locally aggressive nature of this lesion as it rapidly eroded into the lung. Sputum cultures showed growth of E. coli, indicating E. coli infection as a possible etiology of this lesion. Pathology was consistent with inflammatory pseudotumor. PMID:26347780

  13. Effects of short-term inhaled fluticasone on oxidative burst of sputum cells in cystic fibrosis patients.

    PubMed

    Dauletbaev, N; Viel, K; Behr, J; Loitsch, S; Buhl, R; Wagner, T O; Bargon, J

    1999-11-01

    Inhaled corticosteroids have been proposed to decrease pulmonary inflammation in cystic fibrosis (CF). In this study the effects of therapy with inhaled fluticasone on clinical and sputum outcomes (leukocyte count, activity of myeloperoxidase, superoxide anion release) in adult CF patients were investigated in an open label design. Twenty-six stable patients (median+/-SD forced expiratory volume in one second (FEV1) 58.1+/-19.9% pred.) were randomly assigned to the study group (500 microg b.i.d., for three weeks) or the control group (n=14; nonsteroid medication). Sputum samples were obtained during inhalation of hypertonic saline (3%, 20 min), which was found not to alter the investigated sputum parameters. No significant changes in clinical parameters, sputum leukocyte count, activity of myeloperoxidase, and baseline superoxide anion release where observed following therapy. Surprisingly, stimulated superoxide anion release increased significantly after therapy (34.1+/-17.7 versus 25.2+/-17.4 nmol x hr(-1) x 10(6) cells, p<0.03) and exceeded spontaneous variability of this parameter (p=0.02 versus control group). In conclusion, in adult cystic fibrosis patients short-term fluticasone therapy had no evident effect on clinical and sputum parameters. Further investigations are necessary to evaluate whether the observed up-regulation of oxidative capacity of inflammatory cells is of concern or benefit in these patients. PMID:10596705

  14. N-acetylcysteine enhances cystic fibrosis sputum penetration and airway gene transfer by highly compacted DNA nanoparticles.

    PubMed

    Suk, Jung Soo; Boylan, Nicholas J; Trehan, Kanika; Tang, Benjamin C; Schneider, Craig S; Lin, Jung-Ming G; Boyle, Michael P; Zeitlin, Pamela L; Lai, Samuel K; Cooper, Mark J; Hanes, Justin

    2011-11-01

    For effective airway gene therapy of cystic fibrosis (CF), inhaled gene carriers must first penetrate the hyperviscoelastic sputum covering the epithelium. Whether clinically studied gene carriers can penetrate CF sputum remains unknown. Here, we measured the diffusion of a clinically tested nonviral gene carrier, composed of poly-l-lysine conjugated with a 10 kDa polyethylene glycol segment (CK(30)PEG(10k)). We found that CK(30)PEG(10k)/DNA nanoparticles were trapped in CF sputum. To improve gene carrier diffusion across sputum, we tested adjuvant regimens consisting of N-acetylcysteine (NAC), recombinant human DNase (rhDNase) or NAC together with rhDNase. While rhDNase alone did not enhance gene carrier diffusion, NAC and NAC + rhDNase increased average effective diffusivities by 6-fold and 13-fold, respectively, leading to markedly greater fractions of gene carriers that may penetrate sputum layers. We further tested the adjuvant effects of NAC in the airways of mice with Pseudomonas aeruginosa lipopolysaccharide (LPS)-induced mucus hypersecretion. Intranasal dosing of NAC prior to CK(30)PEG(10k)/DNA nanoparticles enhanced gene expression by up to ~12-fold compared to saline control, reaching levels observed in the lungs of mice without LPS challenge. Our findings suggest that a promising synthetic nanoparticle gene carrier may transfer genes substantially more effectively to lungs of CF patients if administered following adjuvant mucolytic therapy with NAC or NAC + rhDNase. PMID:21829177

  15. Quantitative fucK gene polymerase chain reaction on sputum and nasopharyngeal secretions to detect Haemophilus influenzae pneumonia.

    PubMed

    Abdeldaim, Guma M K; Strålin, Kristoffer; Olcén, Per; Blomberg, Jonas; Mölling, Paula; Herrmann, Björn

    2013-06-01

    A quantitative polymerase chain reaction (PCR) for the fucK gene was developed for specific detection of Haemophilus influenzae. The method was tested on sputum and nasopharyngeal aspirate (NPA) from 78 patients with community-acquired pneumonia (CAP). With a reference standard of sputum culture and/or serology against the patient's own nasopharyngeal isolate, H. influenzae etiology was detected in 20 patients. Compared with the reference standard, fucK PCR (using the detection limit 10(5) DNA copies/mL) on sputum and NPA showed a sensitivity of 95.0% (19/20) in both cases, and specificities of 87.9% (51/58) and 89.5% (52/58), respectively. In a receiver operating characteristic curve analysis, sputum fucK PCR was found to be significantly superior to sputum P6 PCR for detection of H. influenzae CAP. NPA fucK PCR was positive in 3 of 54 adult controls without respiratory symptoms. In conclusion, quantitative fucK real-time PCR provides a sensitive and specific identification of H. influenzae in respiratory secretions. PMID:23541117

  16. Tissue hyaluronan expression, as reflected in the sputum of lung cancer patients, is an indicator of malignancy.

    PubMed

    Rangel, M P; de Sá, V K; Martins, V; Martins, J R M; Parra, E R; Mendes, A; Andrade, P C; Reis, R M; Longatto-Filho, A; Oliveira, C Z; Takagaki, T; Carraro, D M; Nader, H B; Capelozzi, V L

    2015-06-01

    Hyaluronan (HA) shows promise for detecting cancerous change in pleural effusion and urine. However, there is uncertainty about the localization of HA in tumor tissue and its relationship with different histological types and other components of the extracellular matrix, such as angiogenesis. We evaluated the association between HA and degree of malignancy through expression in lung tumor tissue and sputum. Tumoral tissue had significantly increased HA compared to normal tissue. Strong HA staining intensity associated with cancer cells was significant in squamous cell carcinoma compared to adenocarcinoma and large cell carcinoma. A significant direct association was found between tumors with a high percentage of HA and MVD (microvessel density) in tumoral stroma. Similarly significant was the direct association between N1 tumors and high levels of HA in cancer cells. Cox multivariate analysis showed significant association between better survival and low HA. HA increased in sputum from lung cancer patients compared to cancer-free and healthy volunteers and a significant correlation was found between HA in sputum and HA in cancer tissue. Localization of HA in tumor tissue was related to malignancy and reflected in sputum, making this an emerging factor for an important diagnostic procedure in patients suspected to have lung cancer. Further study in additional patients in a randomized prospective trial is required to finalize these results and to validate our quantitative assessment of HA, as well as to couple it to gold standard sputum cytology.

  17. N-acetylcysteine enhances cystic fibrosis sputum penetration and airway gene transfer by highly compacted DNA nanoparticles.

    PubMed

    Suk, Jung Soo; Boylan, Nicholas J; Trehan, Kanika; Tang, Benjamin C; Schneider, Craig S; Lin, Jung-Ming G; Boyle, Michael P; Zeitlin, Pamela L; Lai, Samuel K; Cooper, Mark J; Hanes, Justin

    2011-11-01

    For effective airway gene therapy of cystic fibrosis (CF), inhaled gene carriers must first penetrate the hyperviscoelastic sputum covering the epithelium. Whether clinically studied gene carriers can penetrate CF sputum remains unknown. Here, we measured the diffusion of a clinically tested nonviral gene carrier, composed of poly-l-lysine conjugated with a 10 kDa polyethylene glycol segment (CK(30)PEG(10k)). We found that CK(30)PEG(10k)/DNA nanoparticles were trapped in CF sputum. To improve gene carrier diffusion across sputum, we tested adjuvant regimens consisting of N-acetylcysteine (NAC), recombinant human DNase (rhDNase) or NAC together with rhDNase. While rhDNase alone did not enhance gene carrier diffusion, NAC and NAC + rhDNase increased average effective diffusivities by 6-fold and 13-fold, respectively, leading to markedly greater fractions of gene carriers that may penetrate sputum layers. We further tested the adjuvant effects of NAC in the airways of mice with Pseudomonas aeruginosa lipopolysaccharide (LPS)-induced mucus hypersecretion. Intranasal dosing of NAC prior to CK(30)PEG(10k)/DNA nanoparticles enhanced gene expression by up to ~12-fold compared to saline control, reaching levels observed in the lungs of mice without LPS challenge. Our findings suggest that a promising synthetic nanoparticle gene carrier may transfer genes substantially more effectively to lungs of CF patients if administered following adjuvant mucolytic therapy with NAC or NAC + rhDNase.

  18. Tissue hyaluronan expression, as reflected in the sputum of lung cancer patients, is an indicator of malignancy

    PubMed Central

    Rangel, M.P.; de Sá, V.K.; Martins, V.; Martins, J.R.M.; Parra, E.R.; Mendes, A.; Andrade, P.C.; Reis, R.M.; Longatto-Filho, A.; Oliveira, C.Z.; Takagaki, T.; Carraro, D.M.; Nader, H.B.; Capelozzi, V.L.

    2015-01-01

    Hyaluronan (HA) shows promise for detecting cancerous change in pleural effusion and urine. However, there is uncertainty about the localization of HA in tumor tissue and its relationship with different histological types and other components of the extracellular matrix, such as angiogenesis. We evaluated the association between HA and degree of malignancy through expression in lung tumor tissue and sputum. Tumoral tissue had significantly increased HA compared to normal tissue. Strong HA staining intensity associated with cancer cells was significant in squamous cell carcinoma compared to adenocarcinoma and large cell carcinoma. A significant direct association was found between tumors with a high percentage of HA and MVD (microvessel density) in tumoral stroma. Similarly significant was the direct association between N1 tumors and high levels of HA in cancer cells. Cox multivariate analysis showed significant association between better survival and low HA. HA increased in sputum from lung cancer patients compared to cancer-free and healthy volunteers and a significant correlation was found between HA in sputum and HA in cancer tissue. Localization of HA in tumor tissue was related to malignancy and reflected in sputum, making this an emerging factor for an important diagnostic procedure in patients suspected to have lung cancer. Further study in additional patients in a randomized prospective trial is required to finalize these results and to validate our quantitative assessment of HA, as well as to couple it to gold standard sputum cytology. PMID:25992645

  19. Antibacterial therapy of aspiration pneumonia in patients with methicillin-resistant Staphylococcus aureus-positive sputum: identification of risk factors.

    PubMed

    Shinoda, Y; Matsuoka, T; Mori, T; Yoshida, S; Ohashi, K; Yoshimura, T; Sugiyama, T

    2016-02-01

    Inappropriate antimicrobial treatment could adversely affect the recovery of patients with aspiration pneumonia. We attempted to identify inappropriate antibacterial treatment and to determine the standard use of anti-methicillin-resistant Staphylococcus aureus (MRSA) drugs in aspiration pneumonia patients with MRSA-positive in sputum. Aspiration pneumonia patients with MRSA-positive sputum treated between January 2013 and May 2013 were included in this study to determine the risk factors for death during hospitalization. The relationship between anti-MRSA medicine use and death during hospitalization was also investigated. More than 10⁷ MRSA colony-forming units in sputum culture, creatinine clearance of less than 30 mL/min, and quinolone use were found to be risk factors for death during hospitalization. The death rate during hospitalization was significantly lower in cases a Geckler classification of 4 or 5 when anti-MRSA treatment was initiated soon after the culture was obtained. Therefore, we concluded that the use of quinolones as antibacterial treatment in aspiration pneumonia patients with MRSA-positive sputum should be avoided and that anti-MRSA treatment should be started in cases with good quality sputum cultures.

  20. Loring AFB, Caribou, Maine. Revised Uniform Summary of Surface Weather Observations (RUSSWO). Parts a-f. Final report

    SciTech Connect

    Not Available

    1980-06-27

    This report is a six-part statistical summary of surface weather observations for Loring AFB, Caribou, ME. It contains the following parts: (A) Weather Conditions; Atmospheric Phenomena; (B) Precipitation, Snowfall and Snow Depth (daily amounts and extreme values); (C) Surface winds; (D) Ceiling versus Visibility; Sky Cover; (E) Psychrometric Summaries (daily maximum and minimum temperatures, extreme maximum and minimum temperatures, psychrometric summary of wet-bulb temperature depression versus dry-bulb temperature, means and standard deviations of dry-bulb, wet-bulb and dew point temperatures and relative humidity); and (F) Pressure Summary (means, standard deviations, and observation counts of station pressure and sea-level pressure). Data in this report are presented in tabular form, in most cases in percentage frequency of occurrence or cumulative percentage frequency of occurrence tables. (Author)

  1. Effect of Nitrite and Nitrate Concentrations on the Performance of AFB-MFC Enriched with High-Strength Synthetic Wastewater.

    PubMed

    Huang, Jian-Sheng; Yang, Ping; Li, Chong-Ming; Guo, Yong; Lai, Bo; Wang, Ye; Feng, Li; Zhang, Yun

    2015-01-01

    In order to study the effect of nitrite and nitrate on the performance of microbial fuel cell, a system combining an anaerobic fluidized bed (AFB) and a microbial fuel cell (MFC) was employed for high-strength nitrogen-containing synthetic wastewater treatment. Before this study, the AFB-MFC had been used to treat high-strength organic wastewater for about one year in a continuous flow mode. The results showed that when the concentrations of nitrite nitrogen and nitrate nitrogen were increased from 1700 mg/L to 4045 mg/L and 545 mg/L to 1427 mg/L, respectively, the nitrite nitrogen and nitrate nitrogen removal efficiencies were both above 99%; the COD removal efficiency went up from 60.00% to 88.95%; the voltage was about 375 ± 15 mV while the power density was at 70 ± 5 mW/m(2). However, when the concentrations of nitrite nitrogen and nitrate nitrogen were above 4045 mg/L and 1427 mg/L, respectively, the removal of nitrite nitrogen, nitrate nitrogen, COD, voltage, and power density were decreased to be 86%, 88%, 77%, 180 mV, and 17 mW/m(2) when nitrite nitrogen and nitrate nitrogen were increased to 4265 mg/L and 1661 mg/L. In addition, the composition of biogas generated in the anode chamber was analyzed by a gas chromatograph. Nitrogen gas, methane, and carbon dioxide were obtained. The results indicated that denitrification happened in anode chamber. PMID:26495144

  2. Effect of Nitrite and Nitrate Concentrations on the Performance of AFB-MFC Enriched with High-Strength Synthetic Wastewater

    PubMed Central

    Huang, Jian-sheng; Yang, Ping; Li, Chong-ming; Guo, Yong; Lai, Bo; Wang, Ye; Feng, Li; Zhang, Yun

    2015-01-01

    In order to study the effect of nitrite and nitrate on the performance of microbial fuel cell, a system combining an anaerobic fluidized bed (AFB) and a microbial fuel cell (MFC) was employed for high-strength nitrogen-containing synthetic wastewater treatment. Before this study, the AFB-MFC had been used to treat high-strength organic wastewater for about one year in a continuous flow mode. The results showed that when the concentrations of nitrite nitrogen and nitrate nitrogen were increased from 1700 mg/L to 4045 mg/L and 545 mg/L to 1427 mg/L, respectively, the nitrite nitrogen and nitrate nitrogen removal efficiencies were both above 99%; the COD removal efficiency went up from 60.00% to 88.95%; the voltage was about 375 ± 15 mV while the power density was at 70 ± 5 mW/m2. However, when the concentrations of nitrite nitrogen and nitrate nitrogen were above 4045 mg/L and 1427 mg/L, respectively, the removal of nitrite nitrogen, nitrate nitrogen, COD, voltage, and power density were decreased to be 86%, 88%, 77%, 180 mV, and 17 mW/m2 when nitrite nitrogen and nitrate nitrogen were increased to 4265 mg/L and 1661 mg/L. In addition, the composition of biogas generated in the anode chamber was analyzed by a gas chromatograph. Nitrogen gas, methane, and carbon dioxide were obtained. The results indicated that denitrification happened in anode chamber. PMID:26495144

  3. Decoupling directed and passive motion in dynamic systems: particle tracking microrheology of sputum.

    PubMed

    Fong, Erika J; Sharma, Yasha; Fallica, Brian; Tierney, Dylan B; Fortune, Sarah M; Zaman, Muhammad H

    2013-04-01

    Probing the physical properties of heterogeneous materials is essential to understand the structure, function and dynamics of complex fluids including cells, mucus, and polymer solutions. Particle tracking microrheology is a useful method to passively probe viscoelastic properties on micron length scales by tracking the thermal motion of beads embedded in the sample. However, errors associated with active motion have limited the implementation to dynamic systems. We present a simple method to decouple active and Brownian motion, enabling particle tracking to be applied to fluctuating heterogeneous systems. We use the movement perpendicular to the major axis of motion in time to calculate rheological properties. Through simulated data we demonstrate that this method removes directed motion and performs equally well when there is no directed motion, with an average percent error of <1%. We use this method to measure glycerol-water mixtures to show the capability to measure a range of materials. Finally, we use this technique to characterize the compliance of human sputum. We also investigate the effect of a liquefaction agent used to prepare sputum for diagnostic purposes. Our results suggest that the addition of high concentration sodium hydroxide increases sample heterogeneity by increasing the maximum observed creep compliance. PMID:23271563

  4. Classification of Mycobacterium tuberculosis in Images of ZN-Stained Sputum Smears

    PubMed Central

    Khutlang, Rethabile; Krishnan, Sriram; Dendere, Ronald; Whitelaw, Andrew; Veropoulos, Konstantinos; Learmonth, Genevieve; Douglas, Tania S.

    2010-01-01

    Screening for tuberculosis (TB) in low- and middle-income countries is centered on the microscope. We present methods for the automated identification of Mycobacterium tuberculosis in images of Ziehl–Neelsen (ZN) stained sputum smears obtained using a bright-field microscope. We segment candidate bacillus objects using a combination of two-class pixel classifiers. The algorithm produces results that agree well with manual segmentations, as judged by the Hausdorff distance and the modified Williams index. The extraction of geometric-transformation-invariant features and optimization of the feature set by feature subset selection and Fisher transformation follow. Finally, different two-class object classifiers are compared. The sensitivity and specificity of all tested classifiers is above 95% for the identification of bacillus objects represented by Fisher-transformed features. Our results may be used to reduce technician involvement in screening for TB, and would be particularly useful in laboratories in countries with a high burden of TB, where, typically, ZN rather than auramine staining of sputum smears is the method of choice. PMID:19726269

  5. Decoupling directed and passive motion in dynamic systems: particle tracking microrheology of sputum

    PubMed Central

    Fong, Erika J.; Sharma, Yasha; Fallica, Brian; Tierney, Dylan B.; Fortune, Sarah M.; Zaman, Muhammad H.

    2013-01-01

    Probing the physical properties of heterogeneous materials is essential to understand the structure, function and dynamics of complex fluids including cells, mucus, and polymer solutions. Particle tracking microrheology is a useful method to passively probe viscoelastic properties on micron length scales by tracking the thermal motion of beads embedded in the sample. However, errors associated with active motion have limited the implementation to dynamic systems. We present a simple method to decouple active and Brownian motion, enabling particle tracking to be applied to fluctuating heterogeneous systems. We use the movement perpendicular to the major axis of motion in time to calculate rheological properties. Through simulated data we demonstrate that this method removes directed motion and performs equally well when there is no directed motion, with an average percent error of <1%. We use this method to measure glycerol-water mixtures to show the capability to measure a range of materials. Finally, we use this technique to characterize the compliance of human sputum. We also investigate the effect of a liquefaction agent used to prepare sputum for diagnostic purposes. Our results suggest that the addition of high concentration sodium hydroxide increases sample heterogeneity by increasing the maximum observed creep compliance. PMID:23271563

  6. Pediatric Cystic Fibrosis Sputum Can Be Chemically Dynamic, Anoxic, and Extremely Reduced Due to Hydrogen Sulfide Formation

    PubMed Central

    Cowley, Elise S.; Kopf, Sebastian H.; LaRiviere, Alejandro

    2015-01-01

    ABSTRACT Severe and persistent bacterial lung infections characterize cystic fibrosis (CF). While several studies have documented the microbial diversity within CF lung mucus, we know much less about the inorganic chemistry that constrains microbial metabolic processes and their distribution. We hypothesized that sputum is chemically heterogeneous both within and between patients. To test this, we measured microprofiles of oxygen and sulfide concentrations as well as pH and oxidation-reduction potentials in 48 sputum samples from 22 pediatric patients with CF. Inorganic ions were measured in 20 samples from 12 patients. In all cases, oxygen was depleted within the first few millimeters below the sputum-air interface. Apart from this steep oxycline, anoxia dominated the sputum environment. Different sputum samples exhibited a broad range of redox conditions, with either oxidizing (16 mV to 355 mV) or reducing (−300 to −107 mV) potentials. The majority of reduced samples contained hydrogen sulfide and had a low pH (2.9 to 6.5). Sulfide concentrations increased at a rate of 0.30 µM H2S/min. Nitrous oxide was detected in only one sample that also contained sulfide. Microenvironmental variability was observed both within a single patient over time and between patients. Modeling oxygen dynamics within CF mucus plugs indicates that anoxic zones vary as a function of bacterial load and mucus thickness and can occupy a significant portion of the mucus volume. Thus, aerobic respiration accounts only partially for pathogen survival in CF sputum, motivating research to identify mechanisms of survival under conditions that span fluctuating redox states, including sulfidic environments. PMID:26220964

  7. Incubation time of Mycobacterium tuberculosis complex sputum cultures in BACTEC MGIT 960: 4weeks of negative culture is enough for physicians to consider alternative diagnoses.

    PubMed

    Ogwang, Sam; Mubiri, Paul; Bark, Charles M; Joloba, Moses L; Boom, W Henry; Johnson, John L

    2015-10-01

    We retrospectively analyzed time to detection of 3747 positive MGIT sputum cultures at a laboratory in a country with heavy burden of tuberculosis. Ninety-nine percent of diagnostic cultures turned positive within 28days, suggesting that physicians may consider alternative diagnoses if sputum cultures remain negative after 4weeks of incubation.

  8. Human sputum cathepsin B degrades proteoglycan, is inhibited by alpha 2-macroglobulin and is modulated by neutrophil elastase cleavage of cathepsin B precursor and cystatin C.

    PubMed Central

    Buttle, D J; Abrahamson, M; Burnett, D; Mort, J S; Barrett, A J; Dando, P M; Hill, S L

    1991-01-01

    The high-Mr alkali-stable form of cathepsin B was purified from purulent human sputum. It was shown to solubilize proteoglycan monomer entrapped in polyacrylamide at a rate comparable with that of human lysosomal cathepsin B. Like the enzyme from lysosomes, sputum cathepsin B was bound by human alpha 2-macroglobulin, which inhibited its action on proteoglycan. Cystatin C in purulent sputum was shown to be the N-terminally truncated form generated by neutrophil elastase cleavage, and sputum cathepsin B was only weakly inhibited by recombinant cystatin C that had been cleaved by neutrophil elastase in vitro. Addition of neutrophil elastase to mucoid sputum led to a 5-fold increase in cathepsin B activity concomitant with a lowering in Mr of the cysteine proteinase from 40,000 to 37,000, i.e. the size of the active enzyme purified from purulent sputum. It is concluded that the high-Mr form of cathepsin B present in purulent sputum is a functional proteinase, unlike similar forms of the enzyme secreted by mammary gland in organ culture. The activity of cathepsin B in sputum is modulated by neutrophil elastase, by a combination of inhibitor inactivation and zymogen activation. Images Fig. 3. Fig. 4. Fig. 5. PMID:1710889

  9. Immunological status of the progeny of breeder hens kept on ochratoxin A (OTA)- and aflatoxin B(1) (AFB(1))-contaminated feeds.

    PubMed

    Ul-Hassan, Zahoor; Khan, Muhammad Zargham; Khan, Ahrar; Javed, Ijaz

    2012-01-01

    This study aimed to evaluate the immunological status of progeny of hens kept on ochratoxin A (OTA)- and aflatoxin B(1) (AFB(1))-contaminated feed. For this purpose, White Leghorn (WL) layer breeder hens (40-weeks-of-age) were divided into six groups (A-F). Hens in Group A were fed a commercial layer ration while those in Groups B and C were kept on a diet amended with 3 and 5 mg OTA/Kg, respectively. Group D was fed a ration containing 5 mg AFB(1)/Kg, while hens in Groups E and F were kept on feed amended with OTA and AFB(1) each. All feedings were for 1, 2, or 3 weeks. Fertile eggs were set for hatching on a weekly basis to obtain progeny of each week separately. At 14 days-of-age, subsets of progeny were euthanized and the frequency of immunoglobulin(s)-bearing cells in their spleen and bursa of Fabricius assessed; at 16 days-of-age, other chicks in each set were utilized to determine their lymphoblastogenic responses against phytohemagglutinin (PHA-P). At 30 days-of-age, the final sub-set of chicks/group was euthanized and their peritoneal macrophages harvested for measurements of phagocytic potential and nitrite production. Relative weights of the bursa of Fabricius and of the spleen were significantly lower in the progeny of hens fed mycotoxin-contaminated diets for 14 and 21 days. The frequencies of IgA-, IgG-, and IgM-bearing cells were also significantly lower in the bursa of Fabricius and spleen of progeny chicks obtained from hens fed the OTA + AFB(1) mixed diet. Feeding contaminated diets to breeder hens also resulted in significantly lower responses to PHA-P. In addition, the percentages of peritoneal macrophages displaying phagocytosis of sheep red blood cells (SRBC), the number of SRBC/macrophage, and nitrite production were each significantly lower in cells from progeny chicks from OTA- and AFB(1)-fed hens. The findings of the present study indicated there were severe immunosuppressive effects in progeny chicks as a result of exposure of their

  10. Early stationary phase culture supernatant accelerates growth of sputum cultures collected after initiation of anti-tuberculosis treatment.

    PubMed

    Kolwijck, E; Friedrich, S O; Karinja, M N; van Ingen, J; Warren, R M; Diacon, A H

    2014-07-01

    We investigated the effect of Mycobacterium tuberculosis culture supernatant added to sputum cultures collected during the first 8 weeks of anti-tuberculosis treatment. With ongoing treatment duration, time to culture positivity decreased significantly in supernatant-enriched cultures, possibly due to stimulation of dormant or slowly metabolizing M. tuberculosis cells.

  11. Early Lung Cancer Detection in Uranium Miners with Abnormal Sputum Cytology

    SciTech Connect

    Saccomanno, G.

    2000-06-30

    ''Early Lung Cancer Detection in Uranium Miners with Abnormal Sputum Cytology'' was funded by the Department of Energy to monitor the health effects of radon exposure and/or cigarette smoke on uranium workers from the Colorado Plateau. The resulting Saccomanno Uranium Workers Archive and data base has been used as a source of information to prove eligibility for compensation under the Radiation Exposure Compensation Act and as the source of primary data tissue for a subcontract and other collaborations with outside investigators. The latter includes a study of radon exposure and lung cancer risk in a non-smoking cohort of uranium miners (subcontract); a study of genetic markers for lung cancer susceptibility; and a study of {sup 210}Pb accumulation in the skull as a biomarker of radon exposure.

  12. Segmentation of touching mycobacterium tuberculosis from Ziehl-Neelsen stained sputum smear images

    NASA Astrophysics Data System (ADS)

    Xu, Chao; Zhou, Dongxiang; Liu, Yunhui

    2015-12-01

    Touching Mycobacterium tuberculosis objects in the Ziehl-Neelsen stained sputum smear images present different shapes and invisible boundaries in the adhesion areas, which increases the difficulty in objects recognition and counting. In this paper, we present a segmentation method of combining the hierarchy tree analysis with gradient vector flow snake to address this problem. The skeletons of the objects are used for structure analysis based on the hierarchy tree. The gradient vector flow snake is used to estimate the object edge. Experimental results show that the single objects composing the touching objects are successfully segmented by the proposed method. This work will improve the accuracy and practicability of the computer-aided diagnosis of tuberculosis.

  13. Asbestos bodies in the sputum of workers exposed to environmental pollution.

    PubMed

    Alderisio, M; Giovagnoli, M R; Cenci, M; Vecchione, A

    1996-01-01

    The diseases related to asbestos exposure (pulmonary fibrosis, broncogenic carcinoma and mesothelioma) are of widespread interest and involve different socio-economic groups of subjects. Since these pathologies have a wide diffusion in the industrial world, we carried out an investigation on two populations occupationally exposed to air pollution and asbestos fibre inhalation (164 traffic policemen of the municipal district of Rome and 218 railwaymen) and on a control group (119 residents in a rural district of Perugia) for the detection of asbestos bodies in the sputum. The results obtained from traffic policemen and railwaymen workers differred significantly from those of the control group. The presence of asbestos bodies in traffic policemen seems to be determined by a strong synergetic effect between gaseous urban pollution, cigarette smoking habits and asbestos dust arising from car brakes and building materials, whereas, in railwaymen it seems to be more directly correlated to exposure.

  14. Early Detection of Lung Cancer with Meso Tetra (4-Carboxyphenyl) Porphyrin-Labeled Sputum

    PubMed Central

    Patriquin, Lara; Merrick, Daniel T.; Hill, David; Holcomb, Richard G.; Lemieux, Madeleine E.; Bennett, Gordon; Karia, Bijal; Rebel, Vivienne I.; Bauer, Thomas

    2016-01-01

    Introduction Early detection of lung cancer in high-risk individuals reduces mortality. Low-dose spiral computed tomography (LDCT) is the current standard but suffers from an exceedingly high false-positive rate (>96%) leading to unnecessary and potentially dangerous procedures. We, therefore, set out to develop a simple, noninvasive, and quantitative assay to detect lung cancer. Methods This proof-of-concept study evaluated the sensitivity/specificity of the CyPath Early Lung Cancer Detection Assay to correctly classify LDCT-confirmed cohorts of high-risk control (n = 102) and cancer (n = 26) subjects. Fluorescence intensity parameters of red fluorescent cells (RFCs) from tetra (4-carboxyphe-nyl) porphyrin (TCPP)-labeled lung sputum samples and subjects’ baseline characteristics were assessed for their predictive power by multivariable logistic regression. A receiver operating characteristic curve was constructed to evaluate the sensitivity/specificity of the CyPath assay. Results RFCs were detectable in cancer subjects more often than in high-risk ones (p = 0.015), and their characteristics differed between cohorts. Two independent predictors of cancer were the mean of RFC average fluorescence intensity/area per subject (p < 0.001) and years smoked (p = 0.003). The CyPath-based classifier had an overall accuracy of 81% in the test population; false-positive rate of 40% and negative predictive value of 83%. Conclusions The tetra (4-carboxyphenyl) porphyrin-based CyPath assay correctly classified study participants into cancer or high-risk cohorts with considerable accuracy. Optimizing sputum collection, sample reading, and refining the classifier should improve sensitivity and specificity. The CyPath assay thus has the potential to complement LDCT screening or serve as a stand-alone approach for early lung cancer detection. PMID:26200451

  15. Improved cultural detection of Burkholderia cepacia from sputum in patients with cystic fibrosis

    PubMed Central

    Wright, R; Moore, J; Shaw, A; Dunbar, K; Dodd, M; Webb, K; Redmond, A; Crowe, M; Murphy, P; Peacock, S; Elborn, J

    2001-01-01

    Aims—To evaluate the sensitivity and specificity of two selective media for the isolation of Burkholderia cepacia from sputum specimens in patients with cystic fibrosis (CF). Methods—In total, 149 expectorated sputum specimens from 113 patients with CF (32 cepacia colonised patients and 81 non-cepacia colonised patients) attending three CF centres were examined for the presence of B cepacia on two selective media: (1) MAST selective agar, a commercially available selective medium widely used in the UK and (2) BCSA (B cepacia selective agar), a new medium recently described, which is used predominantly in North America. Results—Burkholderia cepacia was isolated from 53 of 149 (35.6%) specimens examined, representing 32 of 113 (28.3%) patients, using both the MAST and BCSA media. Growth was most rapid on BCSA with all (53 of 53) isolates detectable after 48 hours, compared with 50 of the 53 isolates on MAST agar, with the remaining three isolates detectable at five days. Twenty eight contaminants were identified on MAST agar and 13 on BCSA agar; mainly Alcaligenes xylosoxidans and yeast on MAST agar and Flavobacterium indologenes on BCSA medium. BCSA was equivalent to MAST agar in its ability to isolate B cepacia from patients with CF with a history of B cepacia infection. Conclusions—The increased selectivity and reduced time to detection of BCSA makes it an attractive alternative to MAST. However, its present limited commercial availability in the UK may delay its use in routine diagnostic laboratories because of complications with media preparation and quality control. Key Words: Burkholderia cepacia • Burkholderia cepacia selective agar • MAST agar • cystic fibrosis PMID:11577134

  16. Optimizing outpatient serial sputum colony counting for studies of tuberculosis treatment in resource-poor settings.

    PubMed

    Sloan, Derek J; Corbett, Elizabeth L; Butterworth, Anthony E; Mwandumba, Henry C; Khoo, Saye H; Mdolo, Aaron; Shani, Doris; Kamdolozi, Mercy; Allen, Jenny; Mitchison, Denis A; Coleman, David J; Davies, Geraint R

    2012-07-01

    Serial Sputum Colony Counting (SSCC) is an important technique in clinical trials of new treatments for tuberculosis (TB). Quantitative cultures on selective Middlebrook agar are used to calculate the rate of bacillary elimination from sputum collected from patients at different time points during the first 2 months of therapy. However, the procedure can be complicated by high sample contamination rates. This study, conducted in a resource-poor setting in Malawi, assessed the ability of different antifungal drugs in selective agar to reduce contamination. Overall, 229 samples were studied and 15% to 27% were contaminated. Fungal organisms were particularly implicated, and samples collected later in treatment were at particular risk (P < 0.001). Amphotericin B (AmB) is the standard antifungal drug used on SSCC plates at a concentration of 10 mg/ml. On selective Middlebrook 7H10 plates, AmB at 30 mg/ml reduced sample contamination by 17% compared with AmB at 10 mg/ml. The relative risk of contamination using AmB at 10 mg/ml was 1.79 (95% confidence interval [CI], 1.25 to 3.55). On Middlebrook 7H11 plates, a combination of AmB at 10 mg/ml and carbendazim at 50 mg/ml was associated with 10% less contamination than AmB at 30 mg/ml. The relative risk of contamination with AmB at 30 mg/ml was 1.79 (95% CI, 1.01 to 3.17). Improved antifungal activity was accompanied by a small reduction in bacillary counts, but this did not affect modeling of bacillary elimination. In conclusion, a combination of AmB and carbendazim optimized the antifungal activity of selective media for growth of TB. We recommend this method to reduce contamination rates and improve SSCC studies in African countries where the burden of TB is highest.

  17. Shemya AFB, Alaska 1992 IRP field investigation report. Volume 3, Appendixes B, C, and D: Final report

    SciTech Connect

    Not Available

    1993-02-01

    The US Air Force is currently investigating 22 sites on Shemya Air Force Base (AFB) to determine if past spill and disposal activities have caused environmental damage. These investigations are being carried out under the Air Force`s Installation Restoration Program (IRP). Field investigations were performed in 1992 to obtain the information needed to assess what future actions will need to be carried out at each site. The island`s drinking water supply was also investigated. Activities completed at 10 selected sites included surface sampling to determine the lateral extent of contamination, subsurface sampling to determine the vertical extent of contamination, and the installation of well points and monitoring wells to determine the direction of groundwater flow and if the groundwater has been affected by a site. Geophysical surveys were performed at most sites to identify site boundaries and check for the presence of buried metal to be avoided during drilling activities. This report, appendices B, C, and D contains information on the following: geophysical contour maps and profile plots; human health risk assessment; and ecological risk assessment.

  18. Unexploded ordnance detection experiments at extensive fully ground-truthed test sites at Yuma Proving Ground and Eglin AFB

    NASA Astrophysics Data System (ADS)

    DeLuca, Clyde C.; Marinelli, Vincent R.; Ressler, Marc A.; Ton, Tuan T.

    1999-08-01

    The US Army Research Laboratory (ARL), under the sponsorship of the Strategic Environmental Research and Development Program, is conducting experiments to establish and enhance the ability of low-frequency, ultra-wideband synthetic aperture radar (SAR) to detect and discriminate unexploded ordnance (UXO). Preliminary investigations using ARL's BoomSAR - a UWB radar mounted atop a mobile boom lift platform - concluded that the radar image texture and frequency-dependent scattering from mines and mine-like targets could be exploited in the development of automatic target detection algorithms. To support further investigations, ARL established extensive UXO test sites at the US Army Yuma Proving Ground, Arizona, and Eglin AFB, Florida. The soils at both test sties have been characterized in terms of physical, chemical and electromagnetic properties. Precise location, depth, and orientation information was recorded for each of the approximately 500 inert ordnance test targets at each site. This information helps researchers to better understand the phenomenology associated with UXO target scattering and to more accurately evaluate and modify data processing programs. The ultimate goal is to develop innovative automatic target detection algorithms that provide a high probability of detection with an acceptable false-alarm rate under varying environmental conditions and operational scenarios. This paper present details on the design and characterization of the two test sites and some initial results from BoomSAR data collections.

  19. TIR1/AFB-Aux/IAA auxin perception mediates rapid cell wall acidification and growth of Arabidopsis hypocotyls

    PubMed Central

    Fendrych, Matyáš; Leung, Jeffrey; Friml, Jiří

    2016-01-01

    Despite being composed of immobile cells, plants reorient along directional stimuli. The hormone auxin is redistributed in stimulated organs leading to differential growth and bending. Auxin application triggers rapid cell wall acidification and elongation of aerial organs of plants, but the molecular players mediating these effects are still controversial. Here we use genetically-encoded pH and auxin signaling sensors, pharmacological and genetic manipulations available for Arabidopsis etiolated hypocotyls to clarify how auxin is perceived and the downstream growth executed. We show that auxin-induced acidification occurs by local activation of H+-ATPases, which in the context of gravity response is restricted to the lower organ side. This auxin-stimulated acidification and growth require TIR1/AFB-Aux/IAA nuclear auxin perception. In addition, auxin-induced gene transcription and specifically SAUR proteins are crucial downstream mediators of this growth. Our study provides strong experimental support for the acid growth theory and clarified the contribution of the upstream auxin perception mechanisms. DOI: http://dx.doi.org/10.7554/eLife.19048.001 PMID:27627746

  20. Hazardous-waste technical-assistance survey, McChord AFB, Washington. Final report, 22-26 Oct 90

    SciTech Connect

    Albrecht, L.B.

    1991-03-01

    A hazardous waste survey was conducted at McChord AFB, Washington, from 22-26 Oct 90 which addressed hazardous waste management and waste disposal practices, explored opportunities for waste minimization, and determined waste-streams. Recommendations include: (1) Shops using aircraft soap should switch to a milder soap; (2) Consider using a siliceous-based absorbant; (3) Use a contractor who accepts wet batteries or neutralize the acid; (4) Accumulation point managers should maintain a log; (5) Conduct frequent refresher training; (6) Upgrade accumulation sites; (7) Analyze used paint filters; (8) Dispose of anti-freeze in the sanitary sewer; (9) Sample NDI chemicals to determine if hazardous; (10) Update the Waste Analysis Plan; (11) Find a method to recover solvent from the washrack; (12) Entomology needs to comply with FIFRA; (13) Triple-rinse pesticide containers; (14) List all accumulation sites and managers in the hazardous waste management plan; (15) Use an off-the-shelf filtration unit in the waterfall paint booths; (16) Label all hazardous waste drums; (17) Dispose of waste latex paint as municiple waste; (18) Disposal of old hazardous waste drums; and (19) Analyze shop rags from CATM to determine toxicity.

  1. Diabetes Reduces the Rate of Sputum Culture Conversion in Patients With Newly Diagnosed Multidrug-Resistant Tuberculosis.

    PubMed

    Salindri, Argita D; Kipiani, Maia; Kempker, Russell R; Gandhi, Neel R; Darchia, Lasha; Tukvadze, Nestani; Blumberg, Henry M; Magee, Matthew J

    2016-09-01

    Background.  Diabetes is a risk factor for active tuberculosis (TB), but little is known about the relationship between diabetes and multidrug-resistant (MDR) TB. We aimed to assess risk factors for primary MDR TB, including diabetes, and determine whether diabetes reduced the rate of sputum culture conversion among patients with MDR TB. Methods.  From 2011 to 2014, we conducted a cohort study at the National Center for Tuberculosis and Lung Diseases in Tbilisi, Georgia. Adult (≥35 years) patients with primary TB were eligible. Multidrug-resistant TB was defined as resistance to at least rifampicin and isoniazid. Patients with capillary glycosylated hemoglobin (HbA1c) ≥ 6.5% or previous diagnosis were defined to have diabetes. Polytomous regression was used to estimate the association of patient characteristics with drug resistance. Cox regression was used to compare rates of sputum culture conversion in patients with and without diabetes. Results.  Among 318 patients with TB, 268 had drug-susceptibility test (DST) results. Among patients with DST results, 19.4% (52 of 268) had primary MDR TB and 13.4% (36 of 268) had diabetes. In multivariable analyses, diabetes (adjusted odds ratio [aOR], 2.51; 95% confidence interval [CI], 1.00-6.31) and lower socioeconomic status (aOR, 3.51; 95% CI, 1.56-8.20) were associated with primary MDR TB. Among patients with primary MDR TB, 44 (84.6%) converted sputum cultures to negative. The rate of sputum culture conversion was lower among patients with diabetes (adjusted hazard ratio [aHR], 0.34; 95% CI, .13-.87) and among smokers (aHR, 0.16; 95% CI, .04-.61). Conclusions.  We found diabetes was associated with an increased risk of primary MDR TB; both diabetes and smoking were associated with a longer time to sputum culture conversion.

  2. Diabetes Reduces the Rate of Sputum Culture Conversion in Patients With Newly Diagnosed Multidrug-Resistant Tuberculosis.

    PubMed

    Salindri, Argita D; Kipiani, Maia; Kempker, Russell R; Gandhi, Neel R; Darchia, Lasha; Tukvadze, Nestani; Blumberg, Henry M; Magee, Matthew J

    2016-09-01

    Background.  Diabetes is a risk factor for active tuberculosis (TB), but little is known about the relationship between diabetes and multidrug-resistant (MDR) TB. We aimed to assess risk factors for primary MDR TB, including diabetes, and determine whether diabetes reduced the rate of sputum culture conversion among patients with MDR TB. Methods.  From 2011 to 2014, we conducted a cohort study at the National Center for Tuberculosis and Lung Diseases in Tbilisi, Georgia. Adult (≥35 years) patients with primary TB were eligible. Multidrug-resistant TB was defined as resistance to at least rifampicin and isoniazid. Patients with capillary glycosylated hemoglobin (HbA1c) ≥ 6.5% or previous diagnosis were defined to have diabetes. Polytomous regression was used to estimate the association of patient characteristics with drug resistance. Cox regression was used to compare rates of sputum culture conversion in patients with and without diabetes. Results.  Among 318 patients with TB, 268 had drug-susceptibility test (DST) results. Among patients with DST results, 19.4% (52 of 268) had primary MDR TB and 13.4% (36 of 268) had diabetes. In multivariable analyses, diabetes (adjusted odds ratio [aOR], 2.51; 95% confidence interval [CI], 1.00-6.31) and lower socioeconomic status (aOR, 3.51; 95% CI, 1.56-8.20) were associated with primary MDR TB. Among patients with primary MDR TB, 44 (84.6%) converted sputum cultures to negative. The rate of sputum culture conversion was lower among patients with diabetes (adjusted hazard ratio [aHR], 0.34; 95% CI, .13-.87) and among smokers (aHR, 0.16; 95% CI, .04-.61). Conclusions.  We found diabetes was associated with an increased risk of primary MDR TB; both diabetes and smoking were associated with a longer time to sputum culture conversion. PMID:27419188

  3. Improving sensitivity of direct microscopy for detection of acid-fast bacilli in sputum: use of chitin in mucus digestion.

    PubMed

    Farnia, P; Mohammadi, F; Zarifi, Z; Tabatabee, D J; Ganavi, J; Ghazisaeedi, K; Farnia, P K; Gheydi, M; Bahadori, M; Masjedi, M R; Velayati, A A

    2002-02-01

    In order to try to improve the results of direct smear microscopy, we used the mucus-digesting quality of chitin in tuberculosis (TB) laboratories. For this purpose, a total of 430 sputum specimens were processed by the N-acetyl-L-cysteine concentration, sodium hypochlorite (NaOCl) liquefaction, chitin sedimentation, and direct microscopy methods. Then, the smear sensitivity for acid-fast bacillus detection by chitin-treated sputum was compared with the sensitivity of smears prepared by other methods. Our results showed that the chitin solution took less time to completely homogenize the mucoid sputum than did the N-acetyl-L-cysteine and NaOCl methods. The N-acetyl-L-cysteine concentration method demonstrated sensitivity and specificity levels of 83 and 97%, respectively. In comparison, the sensitivity of chitin sedimentation was 80%, with a specificity of 96.7%. The NaOCl liquefaction method showed a sensitivity of 78%, with a specificity of 96%. Finally, the sensitivity of direct microscopy was lower than those of the other tested methods and was only 46%, with a specificity of 90%. The chitin and NaOCl liquefaction methods are both easy to perform, and they do not require additional equipment (centrifuges). Also, our results demonstrated that the chitin method is less time-consuming than the NaOCl method, since only 30 min of incubation is required to bring complete sedimentation of bacilli in chitin-treated sputum whereas the NaOCl method needs 10 to 12 h to give the same results in the same sputum specimens. Therefore, the chitin liquefaction and sedimentation method may provide better results in TB laboratories of developing countries than the N-acetyl-L-cysteine concentration, NaOCl overnight sedimentation, and direct smear microscopy methods.

  4. Diabetes Reduces the Rate of Sputum Culture Conversion in Patients With Newly Diagnosed Multidrug-Resistant Tuberculosis

    PubMed Central

    Salindri, Argita D.; Kipiani, Maia; Kempker, Russell R.; Gandhi, Neel R.; Darchia, Lasha; Tukvadze, Nestani; Blumberg, Henry M.; Magee, Matthew J.

    2016-01-01

    Background. Diabetes is a risk factor for active tuberculosis (TB), but little is known about the relationship between diabetes and multidrug-resistant (MDR) TB. We aimed to assess risk factors for primary MDR TB, including diabetes, and determine whether diabetes reduced the rate of sputum culture conversion among patients with MDR TB. Methods. From 2011 to 2014, we conducted a cohort study at the National Center for Tuberculosis and Lung Diseases in Tbilisi, Georgia. Adult (≥35 years) patients with primary TB were eligible. Multidrug-resistant TB was defined as resistance to at least rifampicin and isoniazid. Patients with capillary glycosylated hemoglobin (HbA1c) ≥ 6.5% or previous diagnosis were defined to have diabetes. Polytomous regression was used to estimate the association of patient characteristics with drug resistance. Cox regression was used to compare rates of sputum culture conversion in patients with and without diabetes. Results. Among 318 patients with TB, 268 had drug-susceptibility test (DST) results. Among patients with DST results, 19.4% (52 of 268) had primary MDR TB and 13.4% (36 of 268) had diabetes. In multivariable analyses, diabetes (adjusted odds ratio [aOR], 2.51; 95% confidence interval [CI], 1.00–6.31) and lower socioeconomic status (aOR, 3.51; 95% CI, 1.56–8.20) were associated with primary MDR TB. Among patients with primary MDR TB, 44 (84.6%) converted sputum cultures to negative. The rate of sputum culture conversion was lower among patients with diabetes (adjusted hazard ratio [aHR], 0.34; 95% CI, .13–.87) and among smokers (aHR, 0.16; 95% CI, .04–.61). Conclusions. We found diabetes was associated with an increased risk of primary MDR TB; both diabetes and smoking were associated with a longer time to sputum culture conversion. PMID:27419188

  5. Effects of combined treatment with rhDNase and airflow oscillations on spinnability of cystic fibrosis sputum in vitro.

    PubMed

    Dasgupta, B; Tomkiewicz, R P; Boyd, W A; Brown, N E; King, M

    1995-08-01

    Treatment with either rhDNase or high-frequency oscillation has been shown to be effective in improving the physical and transport properties of airway secretions in cystic fibrosis (CF). The objects of this in vitro study was to examine whether combined treatment with oscillation and rhDNase results in greater change of CF sputum spinnability than either treatment by itself. Aliquots of sputum (0.4 g) from eight CF patients were subjected to the following protocols for 15 minutes and then followed for a total of 30 minutes: 1) incubation with 0.04 ml DNase 50 micrograms rhDNase/normal saline (10% dilution) at 37 degrees C to achieve 5 micrograms DNase/g of sputum final concentration; 2) airflow oscillation at 27 Hz similar to the airflow magnitude produced by a commercial high-frequency chest compression (HFCC) device; 3) negative control with no treatment; 4) positive (dilution) control, incubating with 10% saline by volume; 5) combination of DNase and oscillation, and 6) combination of saline and oscillation. For each protocol, sputum spinnability (in mm, mean +/- SD) was measured by means of a filancemeter at baseline, 15, and 30 minutes. Treatment with DNase decreased spinnability significantly more than either saline or oscillation at 15 and 30 minutes (P < 0.02 and P < 0.04, respectively). Incubation with saline or oscillation of CF sputum for 15 and 30 minutes decreased spinnability significantly compared with control. The combination of DNase and oscillation decreased spinnability significantly more than treatment with DNase alone (3.74 +/- 0.45 vs. 6.54 +/- 0.73 at 15 minutes, P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

  6. Control efficiency determination of sudden expansion incinerator bldg 348, Kelly AFB, Texas. Final report, 19 July 1995-11 January 1996

    SciTech Connect

    O`Brien, R.J.

    1996-06-01

    Compliance emissions testing and Volatile Organic Compound (VOC) destruction efficiency determination were conducted on the Sudden Expansion (SUE) Incinerator located at the Kelly AFB Fuel Accessory Test Facility, Bldg 348. The purpose of the Kelly AFB SUE Incinerator is to destroy calibration fluid vapors emitted from fuel accessory test stands located in Bldg 348. The incinerator can also be used to destroy liquid waste calibration fluid by burning it as a supplemental fuel. Emissions testing was conducted during combustion of both vapors and liquid calibration fluid. For purposes of determining the incinerator VOC destruction efficiency, monitoring for Total VOC concentration in the inlet air stream was conducted on 19-20 July 1995. Emissions testing of the incinerator exhaust was conducted on 10-11 January 1996 and included monitoring for Total VOC, oxides of nitrogen (NOx), carbon monoxide (CO), and visible emissions.

  7. Pure neuritic leprosy: Resolving diagnostic issues in acid fast bacilli (AFB)-negative nerve biopsies: A single centre experience from South India

    PubMed Central

    Hui, Monalisa; Uppin, Megha S.; Challa, Sundaram; Meena, A. K.; Kaul, Subhash

    2015-01-01

    Background and Purpose: Demonstration of lepra bacilli is essential for definite or unequivocal diagnosis of pure neuritic leprosy (PNL) on nerve biopsy. However, nerves always do not show bacilli owing to the changes of previous therapy or due to low bacillary load in tuberculoid forms. In absence of granuloma or lepra bacilli, other morphologic changes in endoneurium and perineurium can be of help in making a probable diagnosis of PNL and treating the patient with multidrug therapy. Materials and Methods: Forty-six biopsies of PNL were retrospectively reviewed and histologic findings were compared with 25 biopsies of non leprosy neuropathies (NLN) including vasculitic neuropathy and chronic inflammatory demyelinating polyneuropathy (CIDP). The distribution of endoneurial infiltrate and fibrosis, perineurial thickening, and myelin abnormalities were compared between PNL and NLN biopsies and analyzed by Chi-square test. Results: Out of 46 PNL casses, 24 (52.17 %) biopsies were negative for acid fast bacilli (AFB). In these cases, the features which favor a diagnosis of AFB-negative PNL were endoneurial infiltrate (51.1%), endoneurial fibrosis (54.2%), perineurial thickening (70.8%), and reduced number of myelinated nerve fibers (75%). Interpretation and Conclusion: Nerve biopsy is an efficient tool to diagnose PNL and differentiate it from other causes of NLN. In absence of AFB, the diagnosis of PNL is challenging. In this article, we have satisfactorily evaluated the various hisopthological features and found that endoneurial inflammation, dense fibrosis, and reduction in the number of myelinated nerve fibers are strong supportive indicators of PNL regardless of AFB positivity. PMID:26425006

  8. Improving the Sensitivity of the Xpert MTB/RIF Assay on Sputum Pellets by Decreasing the Amount of Added Sample Reagent: a Laboratory and Clinical Evaluation

    PubMed Central

    Amisano, Danielle; Mboowa, Gerald; Ssengooba, Willy; Blakemore, Robert; Kubiak, Rachel W.; Armstrong, Derek T.; Jones, Martin; Manabe, Yukari C.; Joloba, Moses L.; Ellner, Jerrold J.; Dorman, Susan E.; Alland, David

    2015-01-01

    The Xpert MTB/RIF (Xpert) assay permits rapid near-patient detection of Mycobacterium tuberculosis in sputum; however, the test sensitivity remains suboptimal in paucibacillary specimens that are negative for acid-fast bacilli using smear microscopy. Xpert testing includes dilution with sample reagent, and when processed sputum pellets are tested, the recommended sample reagent/pellet ratio is 3:1. We evaluated whether a decreased sample reagent/pellet ratio of 2:1 increased Xpert sensitivity compared to the recommended 3:1. The limit of detection was determined by inoculating serial dilutions of M. tuberculosis into sputum samples, preparing sputum pellets, and testing each pellet by Xpert at both sample reagent ratios. Processed sputum pellets obtained from M. tuberculosis culture-positive clinical specimens were also tested by Xpert at both ratios. Among spiked sputum pellets, the limit of detection was 1,478 CFU/ml (95% confidence interval [CI], 1,211 to 1,943) at a 3:1 ratio and decreased to 832 CFU/ml (95% CI, 671 to 1,134) at 2:1. The proportion of specimens in which M. tuberculosis was detected was greater at 2:1 than at 3:1 for almost all numbers of CFU/ml; this difference was most prominent at lower numbers of CFU/ml. Among 134 concentrated sputum pellets from the clinical study, the sensitivity of Xpert at 2:1 was greater than at 3:1 overall (80% versus 72%; P = 0.03) and for smear-negative specimens (67% versus 58%; P = 0.12). For Xpert testing of sputum pellets, using a lower sample reagent/pellet ratio increased M. tuberculosis detection, especially for paucibacillary specimens. Our study supports use of a 2:1 sample reagent/pellet dilution for Xpert testing of sputum pellets. PMID:25653410

  9. Air toxics and epigenetic effects: ozone altered microRNAs in the sputum of human subjects

    PubMed Central

    Fry, Rebecca C.; Rager, Julia E.; Bauer, Rebecca; Sebastian, Elizabeth; Peden, David B.; Jaspers, Ilona

    2014-01-01

    Ozone (O3) is a criteria air pollutant that is associated with numerous adverse health effects, including altered respiratory immune responses. Despite its deleterious health effects, possible epigenetic mechanisms underlying O3-induced health effects remain understudied. MicroRNAs (miRNAs) are epigenetic regulators of genomic response to environmental insults and unstudied in relationship to O3 inhalation exposure. Our objective was to test whether O3 inhalation exposure significantly alters miRNA expression profiles within the human bronchial airways. Twenty healthy adult human volunteers were exposed to 0.4 ppm O3 for 2 h. Induced sputum samples were collected from each subject 48 h preexposure and 6 h postexposure for evaluation of miRNA expression and markers of inflammation in the airways. Genomewide miRNA expression profiles were evaluated by microarray analysis, and in silico predicted mRNA targets of the O3-responsive miRNAs were identified and validated against previously measured O3-induced changes in mRNA targets. Biological network analysis was performed on the O3-associated miRNAs and mRNA targets to reveal potential associated response signaling and functional enrichment. Expression analysis of the sputum samples revealed that O3 exposure significantly increased the expression levels of 10 miRNAs, namely miR-132, miR-143, miR-145, miR-199a*, miR-199b-5p, miR-222, miR-223, miR-25, miR-424, and miR-582-5p. The miRNAs and their predicted targets were associated with a diverse range of biological functions and disease signatures, noted among them inflammation and immune-related disease. The present study shows that O3 inhalation exposure disrupts select miRNA expression profiles that are associated with inflammatory and immune response signaling. These findings provide novel insight into epigenetic regulation of responses to O3 exposure. PMID:24771714

  10. A NEW AND RAPID METHOD FOR THE ISOLATION AND CULTIVATION OF TUBERCLE BACILLI DIRECTLY FROM THE SPUTUM AND FECES

    PubMed Central

    Petroff, S. A.

    1915-01-01

    Sixty-nine positive cultures were obtained from sixty-nine specimens of sputum from practically all stages of tuberculosis. Six of these specimens were negative by direct microscopic examination, but the cultures gave positive findings. These six specimens have been positive for tubercle bacilli at some time. Nineteen positive cultures were isolated from thirty-two specimens of feces. All these thirty-two specimens, upon direct microscopical examination, gave positive findings, some showing only a few tubercle bacilli. Six specimens were not free from contaminating organisms, and the remaining seven were negative. The method presented in this paper has proved very simple and accurate for the isolation of tubercle bacilli from sputum. The partial success in isolating tubercle bacilli from feces may be due to the fact that many of the bacilli may be dead. PMID:19867850

  11. Results of sputum, cytology among workers exposed to vinyl chloride monomer and to poly(vinyl chloride).

    PubMed

    Maltoni, C; Lodi, P

    1981-10-01

    The results of systematic cytological sputum examinations of 3,380 Italian VC-PVC industry workers and of 2,287 workers in other industries at different potential risk and chosen as control groups are reported. The results indicate an increase in cellular abnormalities and dysplasias in the epithelium of the respiratory tract among VC-PVC workers. These data are in line with experimental results showing that VC produces lung tumors in mice and with early epidemiological evidence among exposed workers.

  12. Development of an Oligonucleotide Array for Direct Detection of Fungi in Sputum Samples from Patients with Cystic Fibrosis▿ †

    PubMed Central

    Bouchara, Jean-Phillippe; Hsieh, Hsin Yi; Croquefer, Sabine; Barton, Richard; Marchais, Veronique; Pihet, Marc; Chang, Tsung Chain

    2009-01-01

    Cystic fibrosis (CF) is the most common inherited genetic disease in Caucasian populations. Besides bacteria, many species of fungi may colonize the respiratory tract of these patients, sometimes leading to true respiratory infections. In this study, an oligonucleotide array capable of identifying 20 fungal species was developed to directly detect fungi in the sputum samples of CF patients. Species-specific oligonucleotide probes were designed from the internal transcribed spacer (ITS) regions of the rRNA operon and immobilized on a nylon membrane. The fungal ITS regions were amplified by PCR and hybridized to the array for species identification. The array was validated by testing 182 target strains (strains which we aimed to identify) and 141 nontarget strains (135 species), and a sensitivity of 100% and a specificity of 99.2% were obtained. The validated array was then used for direct detection of fungi in 57 sputum samples from 39 CF patients, and the results were compared to those obtained by culture. For 16 sputum samples, the results obtained by the array corresponded with those obtained by culture. For 33 samples, the array detected more fungal species than culture did, while the reverse was found for eight samples. The accuracy of the array for fungal detection in sputum samples was confirmed (or partially confirmed) in some samples by cloning and resequencing the amplified ITS fragments. The present array is a useful tool for both the simultaneous detection of multiple fungal species present in the sputa of CF patients and the identification of fungi isolated from these patients. PMID:19020057

  13. Smoking in Asthma Is Associated with Elevated Levels of Corticosteroid Resistant Sputum Cytokines—An Exploratory Study

    PubMed Central

    Spears, Mark; McSharry, Charles; Chaudhuri, Rekha; Weir, Christopher J.; de Wet, Carl; Thomson, Neil C.

    2013-01-01

    Background Current cigarette smoking is associated with reduced acute responses to corticosteroids and worse clinical outcomes in stable chronic asthma. The mechanism by which current smoking promotes this altered behavior is currently unclear. Whilst cytokines can induce corticosteroid insensitivity in-vitro, how current and former smoking affects airway cytokine concentrations and their responses to oral corticosteroids in stable chronic asthma is unclear. Objectives To examine blood and sputum cytokine concentrations in never, ex and current smokers with asthma before and after oral corticosteroids. Methods Exploratory study utilizing two weeks of oral dexamethasone (equivalent to 40 mg/day prednisolone) in 22 current, 21 never and 10 ex-smokers with asthma. Induced sputum supernatant and plasma was obtained before and after oral dexamethasone. 25 cytokines were measured by multiplex microbead system (Invitrogen, UK) on a Luminex platform. Results Smokers with asthma had elevated sputum cytokine interleukin (IL) -6, -7, and -12 concentrations compared to never smokers with asthma. Few sputum cytokine concentrations changed in response to dexamethasone IL-17 and IFNα increased in smokers, CCL4 increased in never smokers and CCL5 and CXCL10 reduced in ex-smokers with asthma. Ex-smokers with asthma appeared to have evidence of an ongoing corticosteroid resistant elevation of cytokines despite smoking cessation. Several plasma cytokines were lower in smokers with asthma compared to never smokers with asthma. Conclusion Cigarette smoking in asthma is associated with a corticosteroid insensitive increase in multiple airway cytokines. Distinct airway cytokine profiles are present in current smokers and never smokers with asthma and could provide an explanatory mechanism for the altered clinical behavior observed in smokers with asthma. PMID:23951170

  14. Orange-Pigmented Sputum as a Manifestation of Smoke Grenade Inhalation Injury.

    PubMed

    Chatzivasiloglou, Fotini; Katsenos, Stamatis; Psara, Anthoula; Tsintiris, Konstantinos

    2016-01-01

    A 34-year-old man presented with scanty hemoptysis, orange-colored expectoration, and mild dyspnea. He was in an enclosed building taking part in a military training exercise inhaling an orange-colored smoke from a smoke grenade ignition. His symptoms developed immediately after the initial exposure but he sought medical assistance 20 hours later because of their persistence. Fiberoptic bronchoscopy was performed revealing diffuse inflammatory tracheobronchial tree with streaky orange-pigmented secretions in the trachea and both main-stem bronchi. Acute tracheobronchitis was diagnosed and the patient was treated with nebulized bronchodilators and intravenous corticosteroids showing complete recovery. To our knowledge, this is the first well-documented report of inhalation injury induced by a smoke bomb explosion including potassium chlorate oxidizer and Sudan I and presenting with orange-pigmented sputum production. Smoke inhalation injury is associated with significant morbidity and mortality. The heterogeneity of the smoke and the large variety of the resulting symptoms may be the reason why a definition, specific diagnostic criteria, and therapeutic guidelines are still lacking.

  15. Clinical Insights from Metagenomic Analysis of Sputum Samples from Patients with Cystic Fibrosis

    PubMed Central

    Evangelista, Jose S.; Schmieder, Robert; Bailey, Barbara; Haynes, Matthew; Furlan, Mike; Maughan, Heather; Edwards, Robert; Rohwer, Forest; Conrad, Douglas

    2014-01-01

    As DNA sequencing becomes faster and cheaper, genomics-based approaches are being explored for their use in personalized diagnoses and treatments. Here, we provide a proof of principle for disease monitoring using personal metagenomic sequencing and traditional clinical microbiology by focusing on three adults with cystic fibrosis (CF). The CF lung is a dynamic environment that hosts a complex ecosystem composed of bacteria, viruses, and fungi that can vary in space and time. Not surprisingly, the microbiome data from the induced sputum samples we collected revealed a significant amount of species diversity not seen in routine clinical laboratory cultures. The relative abundances of several species changed as clinical treatment was altered, enabling the identification of the climax and attack communities that were proposed in an earlier work. All patient microbiomes encoded a diversity of mechanisms to resist antibiotics, consistent with the characteristics of multidrug-resistant microbial communities that are commonly observed in CF patients. The metabolic potentials of these communities differed by the health status and recovery route of each patient. Thus, this pilot study provides an example of how metagenomic data might be used with clinical assessments for the development of treatments tailored to individual patients. PMID:24478471

  16. Metagenome and Metatranscriptome Profiling of Moderate and Severe COPD Sputum in Taiwanese Han Males

    PubMed Central

    Wu, Lawrence Shih-Hsin; Weng, Julia Tzu-Ya

    2016-01-01

    Chronic obstructive pulmonary disease (COPD) is an inflammatory lung disorder characterized by the progressive obstruction of airflow and is currently the fourth leading cause of death in the world. The pathogenesis of COPD is thought to involve bacterial infections and inflammations. Owing to advancement in sequencing technology, evidence is emerging that supports an association between the lung microbiome and COPD. However, few studies have looked into the expression profile of the bacterial communities in the COPD lungs. In this study, we analyzed the sputum microbiome of four moderate and four severe COPD male patients both at the DNA and RNA level, using next generation sequencing technology. We found that bacterial composition determined by 16S rRNA gene sequencing may not directly translate to the set of actively expressing bacteria as defined by transcriptome sequencing. The two sequencing data agreed on Prevotella, Rothia, Neisseria, Porphyromonas, Veillonella, Fusobacterium and Streptococcus being among the most differentially abundant genera between the moderate and severe COPD samples, supporting their association with COPD severity. However, the two sequencing analyses disagreed on the relative abundance of these bacteria in the two COPD groups, implicating the importance of studying the actively expressing bacteria for enriching our understanding of COPD. Though we have described the metatranscriptome profiles of the lung microbiome in moderate and severe COPD, further investigations are required to determine the functional basis underlying the relationship between the microbial species in the lungs and pathogenesis of COPD. PMID:27428540

  17. Trace incorporation of heavy water reveals slow and heterogeneous pathogen growth rates in cystic fibrosis sputum

    PubMed Central

    Kopf, Sebastian H.; Sessions, Alex L.; Cowley, Elise S.; Reyes, Carmen; Van Sambeek, Lindsey; Hu, Yang; Orphan, Victoria J.; Kato, Roberta; Newman, Dianne K.

    2016-01-01

    Effective treatment for chronic infections is undermined by a significant gap in understanding of the physiological state of pathogens at the site of infection. Chronic pulmonary infections are responsible for the morbidity and mortality of millions of immunocompromised individuals worldwide, yet drugs that are successful in laboratory culture are far less effective against pathogen populations persisting in vivo. Laboratory models, upon which preclinical development of new drugs is based, can only replicate host conditions when we understand the metabolic state of the pathogens and the degree of heterogeneity within the population. In this study, we measured the anabolic activity of the pathogen Staphylococcus aureus directly in the sputum of pediatric patients with cystic fibrosis (CF), by combining the high sensitivity of isotope ratio mass spectrometry with a heavy water labeling approach to capture the full range of in situ growth rates. Our results reveal S. aureus generation times with a median of 2.1 d, with extensive growth rate heterogeneity at the single-cell level. These growth rates are far below the detection limit of previous estimates of CF pathogen growth rates, and the rates are slowest in acutely sick patients undergoing pulmonary exacerbations; nevertheless, they are accessible to experimental replication within laboratory models. Treatment regimens that include specific antibiotics (vancomycin, piperacillin/tazobactam, tobramycin) further appear to correlate with slow growth of S. aureus on average, but follow-up longitudinal studies must be performed to determine whether this effect holds for individual patients. PMID:26715741

  18. Trace incorporation of heavy water reveals slow and heterogeneous pathogen growth rates in cystic fibrosis sputum

    NASA Astrophysics Data System (ADS)

    Kopf, Sebastian H.; Sessions, Alex L.; Cowley, Elise S.; Reyes, Carmen; Van Sambeek, Lindsey; Hu, Yang; Orphan, Victoria J.; Kato, Roberta; Newman, Dianne K.

    2016-01-01

    Effective treatment for chronic infections is undermined by a significant gap in understanding of the physiological state of pathogens at the site of infection. Chronic pulmonary infections are responsible for the morbidity and mortality of millions of immunocompromised individuals worldwide, yet drugs that are successful in laboratory culture are far less effective against pathogen populations persisting in vivo. Laboratory models, upon which preclinical development of new drugs is based, can only replicate host conditions when we understand the metabolic state of the pathogens and the degree of heterogeneity within the population. In this study, we measured the anabolic activity of the pathogen Staphylococcus aureus directly in the sputum of pediatric patients with cystic fibrosis (CF), by combining the high sensitivity of isotope ratio mass spectrometry with a heavy water labeling approach to capture the full range of in situ growth rates. Our results reveal S. aureus generation times with a median of 2.1 d, with extensive growth rate heterogeneity at the single-cell level. These growth rates are far below the detection limit of previous estimates of CF pathogen growth rates, and the rates are slowest in acutely sick patients undergoing pulmonary exacerbations; nevertheless, they are accessible to experimental replication within laboratory models. Treatment regimens that include specific antibiotics (vancomycin, piperacillin/tazobactam, tobramycin) further appear to correlate with slow growth of S. aureus on average, but follow-up longitudinal studies must be performed to determine whether this effect holds for individual patients.

  19. Induced sputum as a method for detection of systemic sclerosis-related interstitial lung disease.

    PubMed

    Yilmaz, Neslihan; Abul, Yasin; Bicakcigil, Muge; Golabi, Pejman; Celikel, Cigdem; Karakurt, Sait; Yavuz, Sule

    2012-07-01

    Inducted sputum (IS) is a non-invasive procedure that can be used for collection of airway secretions. The aim of our study is to evaluate the clinical usefulness of IS for detection of airway inflammation in systemic sclerosis (SSc). Bronchoalveolar lavage and IS were performed to 20 patients with SSc. Eighteen patients who were referred to pulmonary medicine for bronchoalveolar lavage due to other reasons were also recruited for cell counts comparisons. Spirometry, echocardiography and thorax CT (HRCT) imaging were also performed to all patients. Mean macrophage and lymphocyte counts were found to be increased in IS of SSc patients compared with that of control (58.4 ± 14.5% vs. 31.3 ± 16.3%, 30.2 ± 15.4% vs. 15.0 ± 11.5% P < 0.001), whereas mean neutrophil count was lower in the SSc patients (4.1 ± 4.5% vs. 17.2 ± 13.1%, P < 0.05). Significant correlations were noted between BAL and IS findings for macrophage (r = 0.55, P = 0.02) lymphocyte (r = 0.65, P < 0.01) and total cell counts (r = 0.45, P = 0.06). IS is an easy and reliable method for the detection of alveolitis and can be used for early detection of lung involvement in scleroderma.

  20. Pneumococcal serotypes in sputum isolates during acute respiratory illness in Edinburgh.

    PubMed

    Gould, G A; Rhind, G B; Morgan, A D; Williamson, G; Calder, M A

    1987-08-01

    During the years 1978-83 serotyping was carried out on all sputum isolates of pneumococci obtained from patients in the chest wards of the City Hospital, Edinburgh. In 402 patients with acute respiratory illness the peak isolation rates occurred from January to April, and the serotype distribution was similar to that seen in previous UK studies, the commonest types being 3, 6, 9, 19, 23, and 8. The overall mortality rate was 8.7%, the serotype distribution in fatal cases reflecting the distribution of the whole group. The presence of mixed infection, predominantly with Haemophilus influenzae, was associated with a lower mortality rate of 3.5%. Nearly all patients (92%) were either elderly or had a chronic underlying disease and only one death occurred in a patient under 70 years who had no pre-existing disease. Of the pneumococcal serotypes isolated from the 292 patients with chronic chest disease, 82% are included in the new 23 valent pneumococcal vaccine and the efficacy of this needs to be assessed further in high risk patients.

  1. Cyanide levels found in infected cystic fibrosis sputum inhibit airway ciliary function.

    PubMed

    Nair, Chandrika; Shoemark, Amelia; Chan, Mario; Ollosson, Sarah; Dixon, Mellissa; Hogg, Claire; Alton, Eric W F W; Davies, Jane C; Williams, Huw D

    2014-11-01

    We have previously reported cyanide at concentrations of up to 150 μM in the sputum of cystic fibrosis patients infected with Pseudomonas aeruginosa and a negative correlation with lung function. Our aim was to investigate possible mechanisms for this association, focusing on the effect of pathophysiologically relevant cyanide levels on human respiratory cell function. Ciliary beat frequency measurements were performed on nasal brushings and nasal air-liquid interface (ALI) cultures obtained from healthy volunteers and cystic fibrosis patients. Potassium cyanide decreased ciliary beat frequency in healthy nasal brushings (n = 6) after 60 min (150 μM: 47% fall, p<0.0012; 75 μM: 32% fall, p<0.0001). Samples from cystic fibrosis patients (n = 3) showed similar results (150 μM: 55% fall, p = 0.001). Ciliary beat frequency inhibition was not due to loss of cell viability and was reversible. The inhibitory mechanism was independent of ATP levels. KCN also significantly inhibited ciliary beat frequency in ALI cultures, albeit to a lesser extent. Ciliary beat frequency measurements on ALI cultures treated with culture supernatants from P. aeruginosa mutants defective in virulence factor production implicated cyanide as a key component inhibiting the ciliary beat frequency. If cyanide production similarly impairs mucocilliary clearance in vivo, it could explain the link with increased disease severity observed in cystic fibrosis patients with detectable cyanide in their airway.

  2. The dynamic motion of pseudo-sputum mucin solution on a vibrating plate.

    PubMed

    Tarao, Norio; Takagi, Osamu

    2008-01-01

    The motion of pseudo-sputum (20% solution of mucin) on a vibrating plate has been investigated using a function generator and an amplifier. Many finger-like liquid columns have appeared at appropriate frequencies (100-130 Hz) from a droplet of the solution on a vibrating plate. The viscosity of the mucin solution was measured using a capillary viscometer of the Maron-Krieger-Sisko type, and it was shown that the shear-thinning viscosity occurred in the lower frequency region contrary to the fact that the most abnormal viscosity of the rheological dilatancy fluid emerges in higher frequency than the frequency in which it behaves the normal one; the most important mechanical cause of this soliton-like phenomenon or the emergence of finger-like column and wrinkles on the surface of fluid is thought to be the rheological dilatancy (shear-thickening viscosity) of source liquid. In connection with this vibrating liquid column phenomenon, the spectrums of productive cough were also investigated, and it was proved that the effective low frequency vibration, which is a characteristic spectrum of productive cough, is essential for the expectoration and is a similar mechanism to the above-mentioned pug mechanism of the finger-like liquid columns seen on the vibrating plate. PMID:18367821

  3. Changes in sputum cytology, airway inflammation and oxidative stress due to chronic inhalation of biomass smoke during cooking in premenopausal rural Indian women.

    PubMed

    Dutta, Anindita; Roychoudhury, Sanghita; Chowdhury, Saswati; Ray, Manas Ranjan

    2013-06-01

    To perform sputum analysis for verification of pulmonary changes in premenopausal rural Indian women chronically exposed to biomass smoke during cooking.Three consecutive morning sputum samples were collected from 196 women (median age 34 years) cooking with biomass and 149 age-matched control women cooking with cleaner fuel liquefied petroleum gas. Smears made on slides were stained with Papanicolaou and Perl's Prussian blue. Airway oxidative stress was estimated as reactive oxygen species (ROS) generation (by flow cytometry) and superoxide dismutase (SOD) level (by spectrophotometry) in sputum cells. Airway inflammation was measured as sputum levels of interleukin (IL)-6, -8 and tumor necrosis factor- alpha (TNF-α). Particulate matter of diameter less than 10 (PM10) was measured using laser photometer while benzene exposure was monitored by measuring trans, trans-muconic acid (t,t-MA) in urine by HPLC-UV. Compared with control, sputum of biomass users contained more neutrophils, lymphocytes, eosinophils, alveolar macrophages, and showed presence of ciliocytophthoria, Charcot-Leyden crystals, Curschmann's spiral. ROS generation was increased by 2-fold while SOD was depleted by 31% in biomass users. They also had higher sputum levels of IL-6, -8 and TNF-α. Levels of PM10 and t,t-MA were 2.9- and 5.8-times higher in biomass-using women. PM10 and t,t-MA levels were positively associated with cellular changes in the sputum, markers of airway inflammation, and oxidative stress. Cooking with biomass alters sputum cytology, and increases airway inflammation and oxidative stress that might result in further amplification of the tissue damaging cascade in women chronically exposed to biomass smoke.

  4. Effects of Autogenic Drainage on Sputum Recovery and Pulmonary Function in People with Cystic Fibrosis: A Systematic Review

    PubMed Central

    Osterling, Kristin; Gilbert, Robert; Dechman, Gail

    2015-01-01

    ABSTRACT Purpose: To determine the effects of short- and long-term use of autogenic drainage (AD) on pulmonary function and sputum recovery in people with cystic fibrosis (CF). Methods: The authors conducted a systematic review of randomized and quasi-randomized clinical trials in which participants were people with CF who use AD as their sole airway clearance technique. Results: Searches in 4 databases and secondary sources using 5 key terms yielded 735 articles, of which 58 contained the terms autogenic drainage and cystic fibrosis. Ultimately, 4 studies, 2 of which were long term, were included. All measured forced expiratory volume in 1 second (FEV1) and found no change. The long-term studies were underpowered to detect change in FEV1; however, the short-term studies found a clinically significant sputum yield (≥4 g). Conclusion: AD has been shown to produce clinically significant sputum yields in a limited number of investigations. The effect of AD on the function of the pulmonary system remains uncertain, and questions have emerged regarding the appropriateness of FEV1 as a valid measure of airway clearance from peripheral lung regions. Further consideration should be given to the use of FEV1 as a primary measure of the effect of AD. PMID:27504031

  5. Feasibility of using fluorescence in situ hybridization (FISH) to detect early gene changes in sputum cells from uranium miners

    SciTech Connect

    Neft, R.E.; Rogers, J.L.; Belinsky, S.A.

    1995-12-01

    Epidemiological studies have shown that combined exposure to radon progeny and tobacco smoke produce a greater than additive or synergistic increase in lung cancer risk. Lung cancer results from multiple genetic changes over a long period of time. An early change that occurs in lung cancer is trisomy 7 which is found in 50% of non-small cell lung cancer and in the far margins of resected lung tumors. The 80% mortality associated with lung cancer is in part related to the high proportion of patients who present with an advanced, unresectable tumor. Therefore, early detection of patients at risk for tumor development is critical to improve treatment of this disease. Currently, it is difficult to detect lung cancer early while it is still amendable by surgery. Saccomanno, G. has shown that premalignant cytologic changes in sputum cells collected from uranium miners can be detected by a skilled, highly trained cytopathologist. A more objective alternative for identifying premalignant cells in sputum may be to determine whether an early genetic change such as trisomy 7 is present in these cells. Fluorescence in situ hybridization (FISH) can be used to identify cells with trisomy 7. The results of this investigation indicate that FISH may prove to be an accurate, efficient method to test at-risk individuals for genetic alterations in bronchial epithelial cells from sputum.

  6. Evaluation of two line probe assays for rapid detection of Mycobacterium tuberculosis, tuberculosis (TB) drug resistance, and non-TB Mycobacteria in HIV-infected individuals with suspected TB.

    PubMed

    Luetkemeyer, Anne F; Kendall, Michelle A; Wu, Xingye; Lourenço, Maria Cristina; Jentsch, Ute; Swindells, Susan; Qasba, Sarojini S; Sanchez, Jorge; Havlir, Diane V; Grinsztejn, Beatriz; Sanne, Ian M; Firnhaber, Cynthia

    2014-04-01

    Limited performance data from line probe assays (LPAs), nucleic acid tests used for the rapid diagnosis of tuberculosis (TB), nontuberculosis mycobacteria (NTM), and Mycobacterium tuberculosis drug resistance are available for HIV-infected individuals, in whom paucibacillary TB is common. In this study, the strategy of testing sputum with GenoType MTBDRplus (MTBDR-Plus) and GenoType Direct LPA (Direct LPA) was compared to a gold standard of one mycobacterial growth indicator tube (MGIT) liquid culture. HIV-positive (HIV(+)) individuals with suspected TB from southern Africa and South America with <7 days of TB treatment had 1 sputum specimen tested with Direct LPA, MTBDR-Plus LPA, smear microscopy, MGIT, biochemical identification of mycobacterial species, and culture-based drug-susceptibility testing (DST). Of 639 participants, 59.3% were MGIT M. tuberculosis culture positive, of which 276 (72.8%) were acid-fast bacillus (AFB) smear positive. MTBDR-Plus had a sensitivity of 81.0% and a specificity of 100%, with sensitivities of 44.1% in AFB smear-negative versus 94.6% in AFB smear-positive specimens. For specimens that were positive for M. tuberculosis by MTBDR-Plus, the sensitivity and specificity for rifampin resistance were 91.7% and 96.6%, respectively, and for isoniazid (INH) they were 70.6% and 99.1%. The Direct LPA had a sensitivity of 88.4% and a specificity of 94.6% for M. tuberculosis detection, with a sensitivity of 72.5% in smear-negative specimens. Ten of 639 MGIT cultures grew Mycobacterium avium complex or Mycobacterium kansasii, half of which were detected by Direct LPA. Both LPA assays performed well in specimens from HIV-infected individuals, including in AFB smear-negative specimens, with 72.5% sensitivity for M. tuberculosis identification with the Direct LPA and 44.1% sensitivity with MTBDR-Plus. LPAs have a continued role for use in settings where rapid identification of INH resistance and clinically relevant NTM are priorities.

  7. Effects of Fluroquinolones in Newly Diagnosed, Sputum-Positive Tuberculosis Therapy: A Systematic Review and Network Meta-Analysis

    PubMed Central

    Li, Dandan; Wang, Tiansheng; Shen, Su; Cheng, Sheng; Yu, Junxian; Zhang, Yang; Zhang, Chao; Tang, Huilin

    2015-01-01

    Background Tuberculosis is a major public health problem especially in developing countries, the comparative efficacy and safety of fluroquinolones (FQs) for adult patients with newly diagnosed, sputum-positive tuberculosis remains controversial. We aimed to investigate the benefits and risks of FQs-containing (addition/substitution) regimens in this population. Methods A network meta-analysis was performed to compare FQs (C: ciprofloxacin; O: ofloxacin; Lo: levofloxacin; M: moxifloxacin; G: gatifloxacin) addition/substitution regimen with standard HRZE regimen (ie isoniazid, rifampicin, pyrazinamide and ethambutol) in newly diagnosed, sputum-positive tuberculosis. Medline, Embase and Cochrane Central Register of Controlled Trials were systematically searched, randomized trials with duration longer than 8 weeks were included. The primary outcome was week-8 sputum negativity, and secondary outcomes included treatment failure, serious adverse events and death from all cause. Results Twelve studies comprising 6465 participants were included in the network meta-analysis. Löwenstein-Jensen culture method showed that HRZEM (OR 4.96, 95% CI 2.83–8.67), MRZE (OR 1.48, 95% CI 1.19–1.84) and HRZM (OR 1.32, 95% CI 1.08–1.62) had more sputum conversion than HRZE by the eighth week, whereas HRC (OR 0.39, 95% CI 0.19–0.77) and HRZO (OR 0.47, 95% CI 0.24–0.92) were worse than HRZE. Moxifloxacin-containing regimens showed more conversion than HRZE by liquid method at the end of two months. But by the end of treatment, FQs-containing regimens didn’t show superiority than HRZE on treatment failure. There were no significant differences between any regimens on other outcomes like serious adverse events and all-cause death. Conclusion This comprehensive network meta-analysis showed that compared with HRZE, moxifloxacin-containing regimens could significantly increase sputum conversion by the eighth week for patients with newly diagnosed pulmonary tuberculosis while HRC

  8. Defining binding efficiency and specificity of auxins for SCF(TIR1/AFB)-Aux/IAA co-receptor complex formation.

    PubMed

    Lee, Sarah; Sundaram, Shanthy; Armitage, Lynne; Evans, John P; Hawkes, Tim; Kepinski, Stefan; Ferro, Noel; Napier, Richard M

    2014-03-21

    Structure-activity profiles for the phytohormone auxin have been collected for over 70 years, and a number of synthetic auxins are used in agriculture. Auxin classification schemes and binding models followed from understanding auxin structures. However, all of the data came from whole plant bioassays, meaning the output was the integral of many different processes. The discovery of Transport Inhibitor-Response 1 (TIR1) and the Auxin F-Box (AFB) proteins as sites of auxin perception and the role of auxin as molecular glue in the assembly of co-receptor complexes has allowed the development of a definitive quantitative structure-activity relationship for TIR1 and AFB5. Factorial analysis of binding activities offered two uncorrelated factors associated with binding efficiency and binding selectivity. The six maximum-likelihood estimators of Efficiency are changes in the overlap matrixes, inferring that Efficiency is related to the volume of the electronic system. Using the subset of compounds that bound strongly, chemometric analyses based on quantum chemical calculations and similarity and self-similarity indices yielded three classes of Specificity that relate to differential binding. Specificity may not be defined by any one specific atom or position and is influenced by coulomb matrixes, suggesting that it is driven by electrostatic forces. These analyses give the first receptor-specific classification of auxins and indicate that AFB5 is the preferred site for a number of auxinic herbicides by allowing interactions with analogues having van der Waals surfaces larger than that of indole-3-acetic acid. The quality factors are also examined in terms of long-standing models for the mechanism of auxin binding.

  9. Differential downward stream of auxin synthesized at the tip has a key role in gravitropic curvature via TIR1/AFBs-mediated auxin signaling pathways.

    PubMed

    Nishimura, Takeshi; Nakano, Hitomi; Hayashi, Ken-ichiro; Niwa, Chiharu; Koshiba, Tomokazu

    2009-11-01

    Since the early days of Darwin, monocot coleoptiles have been used to investigate indole-3-acetic acid (IAA) production, polar transport and tropisms. Here, using maize coleoptiles, we first showed that polar transport of IAA synthesized at the tip region is regulated by ZmPIN(s). Then, the TIR/AFBs-mediated auxin signaling pathway corresponds to the asymmetric IAA flow after gravi-stimulus, which results in tropic curvature. When [(13)C(11)(15)N(2)]Trp was applied to coleoptile tips, substantial amounts of the stable isotope were incorporated into IAA at the tip region, and the labeled IAA was transported in a polar manner at approximately 7 mm h(-1). Immunohistochemical analyses revealed that ZmPIN1(s) was present in almost all cells. ZmPIN1(s) showed a relatively non-polar distribution at the tip, but a basal cellular localization at lower regions. Application of the IAA transport inhibitors 1-N-naphthylphthalamic acid (NPA) and brefeldin A (BFA) at the very tip region almost completely inhibited IAA movement from the tip. These inhibitors also severely suppressed gravitropic bending. PEO-IAA, an auxin antagonist that binds to TIR1/AFBs, suppressed not only the expression of an auxin-responsive ZmSAUR2 gene, but also gravitropic curvature. Expression of ZmSAUR2 was up-regulated on the lower side and down-regulated on the upper side of the coleoptile elongation zone, corresponding to the asymmetric IAA distribution. These results indicate that the asymmetric downward streams of IAA control the differential growth rate of the cells by attenuating TIR1/AFBs-mediated auxin response genes, including ZmSAUR2, and therefore result in tropic curvature. PMID:19897572

  10. Monitoring and Data Analysis for the Vadose Zone Monitoring System (VZMS), McClellan AFB. Quarterly Status Report (2/20/98 - 5/20/98)

    SciTech Connect

    Zawislanski, P.T.; Mountford, H.S.Monitoring and Data Analysis; for the Vadose Zone Monitoring System; Mountford, H.S.; Dahlquist, R.; Rodriguez, S.J.

    1998-06-18

    This report contains information on field and laboratory work performed between February 20th, 1998 and May 20th, 1998, at site S-7 in IC 34, at McClellan AFB. At this location, a Vadose Zone Monitoring System (VZMS) (LBNL, 1996) is currently being used to collect subsurface data including hydraulic potential, soil gas pressure, moisture content, water chemistry, gas chemistry, and temperature. This report describes: moisture content changes, based on neutron logging; gas-phase VOC concentrations; aqueous-phase VOC concentrations; temperature profiles; and installation of new instrument cluster.

  11. Automated detection of tuberculosis on sputum smeared slides using stepwise classification

    NASA Astrophysics Data System (ADS)

    Divekar, Ajay; Pangilinan, Corina; Coetzee, Gerrit; Sondh, Tarlochan; Lure, Fleming Y. M.; Kennedy, Sean

    2012-03-01

    Routine visual slide screening for identification of tuberculosis (TB) bacilli in stained sputum slides under microscope system is a tedious labor-intensive task and can miss up to 50% of TB. Based on the Shannon cofactor expansion on Boolean function for classification, a stepwise classification (SWC) algorithm is developed to remove different types of false positives, one type at a time, and to increase the detection of TB bacilli at different concentrations. Both bacilli and non-bacilli objects are first analyzed and classified into several different categories including scanty positive, high concentration positive, and several non-bacilli categories: small bright objects, beaded, dim elongated objects, etc. The morphological and contrast features are extracted based on aprior clinical knowledge. The SWC is composed of several individual classifiers. Individual classifier to increase the bacilli counts utilizes an adaptive algorithm based on a microbiologist's statistical heuristic decision process. Individual classifier to reduce false positive is developed through minimization from a binary decision tree to classify different types of true and false positive based on feature vectors. Finally, the detection algorithm is was tested on 102 independent confirmed negative and 74 positive cases. A multi-class task analysis shows high accordance rate for negative, scanty, and high-concentration as 88.24%, 56.00%, and 97.96%, respectively. A binary-class task analysis using a receiver operating characteristics method with the area under the curve (Az) is also utilized to analyze the performance of this detection algorithm, showing the superior detection performance on the high-concentration cases (Az=0.913) and cases mixed with high-concentration and scanty cases (Az=0.878).

  12. 15q12 Variants, Sputum Gene Promoter Hypermethylation, and Lung Cancer Risk: A GWAS in Smokers

    PubMed Central

    Leng, Shuguang; Liu, Yushi; Weissfeld, Joel L.; Thomas, Cynthia L.; Han, Younghun; Picchi, Maria A.; Edlund, Christopher K.; Willink, Randall P.; Gaither Davis, Autumn L.; Do, Kieu C.; Nukui, Tomoko; Zhang, Xiequn; Burki, Elizabeth A.; Van Den Berg, David; Romkes, Marjorie; Gauderman, W. James; Crowell, Richard E.; Tesfaigzi, Yohannes; Stidley, Christine A.; Amos, Christopher I.; Siegfried, Jill M.; Gilliland, Frank D.

    2015-01-01

    Background: Lung cancer is the leading cause of cancer-related mortality worldwide. Detection of promoter hypermethylation of tumor suppressor genes in exfoliated cells from the lung provides an assessment of field cancerization that in turn predicts lung cancer. The identification of genetic determinants for this validated cancer biomarker should provide novel insights into mechanisms underlying epigenetic reprogramming during lung carcinogenesis. Methods: A genome-wide association study using generalized estimating equations and logistic regression models was conducted in two geographically independent smoker cohorts to identify loci affecting the propensity for cancer-related gene methylation that was assessed by a 12-gene panel interrogated in sputum. All statistical tests were two-sided. Results: Two single nucleotide polymorphisms (SNPs) at 15q12 (rs73371737 and rs7179575) that drove gene methylation were discovered and replicated with rs73371737 reaching genome-wide significance (P = 3.3×10–8). A haplotype carrying risk alleles from the two 15q12 SNPs conferred 57% increased risk for gene methylation (P = 2.5×10–9). Rs73371737 reduced GABRB3 expression in lung cells and increased risk for smoking-induced chronic mucous hypersecretion. Furthermore, subjects with variant homozygote of rs73371737 had a two-fold increase in risk for lung cancer (P = .0043). Pathway analysis identified DNA double-strand break repair by homologous recombination (DSBR-HR) as a major pathway affecting susceptibility for gene methylation that was validated by measuring chromatid breaks in lymphocytes challenged by bleomycin. Conclusions: A functional 15q12 variant was identified as a risk factor for gene methylation and lung cancer. The associations could be mediated by GABAergic signaling that drives the smoking-induced mucous cell metaplasia. Our findings also substantiate DSBR-HR as a critical pathway driving epigenetic gene silencing. PMID:25713168

  13. Simplified detection of Mycobacterium tuberculosis in sputum using smear microscopy and PCR with molecular beacons.

    PubMed

    Haldar, Sagarika; Chakravorty, Soumitesh; Bhalla, Manpreet; De Majumdar, Shyamasree; Tyagi, Jaya Sivaswami

    2007-10-01

    The prompt diagnosis of smear-negative cases is a prerequisite to controlling tuberculosis (TB). Several new laboratory approaches, including nucleic acid amplification (NAA), are being evaluated in various disease settings to meet this challenge. However, NAA needs simplification before it is widely accepted. Furthermore, a supporting smear result improves confidence in and reliability of PCR. In this context, an asymmetric devR PCR assay using two molecular beacon probes for visual or fluorimetric end-point detection of Mycobacterium tuberculosis was developed. The assays reproducibly detected 25 fg M. tuberculosis DNA versus 100 fg by conventional gel electrophoresis (henceforth referred to as gel assay). The devR and IS6110 PCR assays were blindly evaluated on sputum specimens obtained from a directly observed-treatment short-course centre. Universal sample processing (USP) smear microscopy and culture were used as a supportive test and the 'gold' standard, respectively. Among the 148 specimens analysed, 120 were M. tuberculosis culture-positive. Amongst the 122 direct smear-negative samples, 96 were culture-positive, of which 61 were detected by USP smear microscopy. All 35 USP smear-negative samples were positive by three or more PCR methods. devR PCR had a sensitivity of 92.5 % in the fluorimetric assay versus 86.7 % by visual inspection and 90.8 % by the gel method. IS6110 PCR performed at almost equivalent levels. devR visual and fluorimetric assays considered together yielded an increased sensitivity of 95 % without compromising on a specificity of 92.9 %. The results suggest that the USP smear test is useful for diagnosing direct smear-negative TB and judiciously restricting PCR testing to only smear-negative samples. When used together, these tests can provide rapid diagnosis of smear-negative TB in a cost-effective manner.

  14. Impact of awareness drives and community-based active tuberculosis case finding in Odisha, India.

    PubMed

    Parija, D; Patra, T K; Kumar, A M V; Swain, B K; Satyanarayana, S; Sreenivas, A; Chadha, V K; Moonan, P K; Oeltmann, J E

    2014-09-01

    India's Revised National Tuberculosis Control programme employs passive case detection. The new sputum smear-positive case detection rate is less than 70% in Odisha State. During April-June 2012, active case finding (ACF) was conducted through awareness drives and field-based tuberculosis (TB) screening in select communities with the lowest case detection rates. During the campaign, 240 sputum smear-positive TB cases were detected. The number of smear-positive cases detected increased by 11% relative to April-June 2011 in intervention communities compared to an 0.8% increase in non-intervention communities. ACF brought TB services closer to the community and increased TB case detection.

  15. Hypertonic saline solutions do not influence the solubility of sputum from secretor and non-secretor cystic fibrosis patients

    PubMed Central

    Barboza, Marcelo A.I.; Brandão de Mattos, Cinara C.; Ferreira, Ana Iara C.; Barja, Paulo R.; Santos de Faria Junior, Newton; de Oliveira, Luís Vicente F.; de Mattos, Luiz C.

    2011-01-01

    Introduction Functional alterations of the cystic fibrosis transmembrane conductance regulator gene (CFTR) increase the viscoelasticity of pulmonary secretions of cystic fibrosis (CF) patients and require the use of therapeutic aerosols. The biochemical properties of exocrine secretions are influenced by the expression of the FUT2 gene which determine the secretor and non-secretor phenotypes of the ABH glycoconjugates. The aim of this study was to determine the influence of secretor and non-secretor phenotypes by means of photoacoustic analysis, both the typical interaction time (t 0) and the solubilization interval (Δt) of the sputum of secretor and non-secretor CF patients nebulized by hypertonic saline solutions at different concentrations. Material and methods Sputum samples were obtained by spontaneous expectoration from 6 secretor and 4 non-secretor patients with CF. Each sample was nebulized with 3%, 6%, and 7% hypertonic saline solutions in a photoacoustic cell. The values of t 0 and Δt were determined using the Origin 7.5® computer program (Microcal Software Inc.). The t-test was employed using the GraphPad Instat 3.0® computer program to calculate the mean and standard deviation for each parameter. Results For all hypertonic saline solutions tested, the mean values of t 0 and Δt do not show statistically significant differences between secretor and non-secretor patients. Conclusions The secretor and non-secretor phenotypes do not influence the in vitro solubilization of the sputum nebulized by hypertonic saline solutions at different concentrations when analysed by photoacoustic technique. PMID:22291775

  16. Simple Detection of the IS6110 Sequence of Mycobacterium tuberculosis Complex in Sputum, Based on PCR with Graphene Oxide.

    PubMed

    Hwang, Sang-Hyun; Kim, Dong-Eun; Sung, Heungsup; Park, Byeong-Min; Cho, Mi-Jeong; Yoon, Ok-Jin; Lee, Do-Hoon

    2015-01-01

    Graphene oxide (GO) has proven to be a satisfactory DNA-sensor platform for applications in enzyme-free signal amplification, fluorescence-based amplification, and nanoparticle-based platforms because of its excellent electrical, thermal, and optical properties. In this study, we designed a novel platform for the fluorescence detection of biomolecules, using a fluorescent dye-labeled primer and GO. We applied this system for the detection of the IS6110 insertion sequence of the Mycobacterium tuberculosis complex (MTB) and evaluated its feasibility for use in molecular diagnostics. Fifty-four sputum specimens were collected at our institution from October 2010 to March 2012. To detect MTB in the samples, we performed PCR amplification of the IS6110 DNA sequence using FAM-labeled primers, after which the PCR amplicon was incubated with GO and the fluorescence was measured. The results were compared with those obtained by conventional real-time quantitative PCR (RQ-PCR). The fluorescence intensity observed increased in a concentration-dependent manner with the FAM-labeled IS6110 amplicon. The results of the PCR-GO system for detecting IS6110 DNA were in good agreement with those obtained with conventional RQ-PCR (kappa statistic = 0.925). The PCR-GO system detected MTB DNA in 23 of 25 RQ-PCR-positive sputum samples (92.0%; 95% CI, 75.0-98.0%), but not in 29 of 29 RQ-PCR-negative sputum samples (100%; 95% CI, 88.1-100.0%). These results indicate the utility of the PCR-GO system in molecular diagnostics.

  17. TB-SA antibody test for diagnosis and monitoring treatment outcome of sputum smear negative pulmonary tuberculosis patients.

    PubMed

    Li, Xinxu; Xu, Hancheng; Jiang, Shiwen; Jing, Kuanhe; Wang, Li; Liu, Xiaoqiu; Li, Weibin; Zhang, Hui; Wang, Lixia

    2011-09-01

    The objectives of this study were to evaluate the suitability of the TB-SA antibody test to diagnose tuberculosis in sputum smear negative (SS-) pulmonary tuberculosis (TB) patients and its applicability for monitoring treatment outcomes in these patients. This study was conducted in three counties/districts in Chongqing Municipality, Liaoning Province, China between June 2005 and June 2007. A total of 432 SS suspected pulmonary TB patients were recruited and their blood was collected prior to treatment, at the end of 1 month of treatment, 2 months of treatment and 6 months of treatment (E6MT). The serum samples were analyzed with a TB-SA antibody test kit. Of the 432 SS suspected pulmonary TB patients, serum samples were obtained at all time points in 316 patients and analyzed. The 316 patients were divided into three groups according to sputum smear and sputum culture results and the chest X-ray results before treatment and at E6MT. Ten point four percent were SS-/culture positive (C+), 73.1% were SS-/culture negative (C-) with X-rays abnormalities, and 16.5% were SS-/C- without X-rays abnormalities. The positive rates for TB-SA antibody in the three groups were 57.6, 44.6 and 44.2%, respectively, before treatment, and 18.2, 19.1 and 26.9%, respectively, at E6MT. There was a significant decrease in TB-SA antibody positivity with treatment for all 3 groups. The TB-SA antibody test may be a useful adjunct to diagnose tuberculosis in SS- pulmonary TB patients, and may be useful for monitoring treatment outcomes of SS- pulmonary TB patients. PMID:22299440

  18. Comparison of Mycobacterial Growth Indicator Tube with Culture on RGM Selective Agar for Detection of Mycobacteria in Sputum Samples from Patients with Cystic Fibrosis.

    PubMed

    Eltringham, Ian; Pickering, Julie; Gough, Helen; Preece, Clair L; Perry, John D

    2016-08-01

    Nontuberculous mycobacteria (NTM) are an important cause of pulmonary disease in patients with cystic fibrosis (CF). A new culture medium (RGM medium) for the isolation of rapidly growing mycobacteria from the sputum of cystic fibrosis patients has recently been reported. The aim of this study was to compare culture of sputum samples on RGM medium with culture using a standard automated liquid culture method. Sputum samples were obtained from 187 distinct patients with CF attending King's College Hospital, London, United Kingdom. Each sample was decontaminated with 3% oxalic acid and inoculated into a mycobacterial growth indicator tube (MGIT) that was monitored for 42 days using the Bactec MGIT 960 instrument. Each sample was also cultured, without decontamination, onto RGM medium, which was incubated for 10 days at 30°C. Mycobacteria were isolated from 28 patients (prevalence, 15%). Mycobacteria were detected in 24 samples (86%) using the MGIT and in 23 samples (82%) using RGM medium (P = 1.00). In this setting, RGM medium showed sensitivity equivalent to that of the MGIT for isolation of NTM from the sputum of patients with CF. RGM medium offers a simple, convenient tool that can be embedded into routine culture methods, allowing the culture of all sputum samples that are submitted from patients with CF.

  19. Comparison of Mycobacterial Growth Indicator Tube with Culture on RGM Selective Agar for Detection of Mycobacteria in Sputum Samples from Patients with Cystic Fibrosis.

    PubMed

    Eltringham, Ian; Pickering, Julie; Gough, Helen; Preece, Clair L; Perry, John D

    2016-08-01

    Nontuberculous mycobacteria (NTM) are an important cause of pulmonary disease in patients with cystic fibrosis (CF). A new culture medium (RGM medium) for the isolation of rapidly growing mycobacteria from the sputum of cystic fibrosis patients has recently been reported. The aim of this study was to compare culture of sputum samples on RGM medium with culture using a standard automated liquid culture method. Sputum samples were obtained from 187 distinct patients with CF attending King's College Hospital, London, United Kingdom. Each sample was decontaminated with 3% oxalic acid and inoculated into a mycobacterial growth indicator tube (MGIT) that was monitored for 42 days using the Bactec MGIT 960 instrument. Each sample was also cultured, without decontamination, onto RGM medium, which was incubated for 10 days at 30°C. Mycobacteria were isolated from 28 patients (prevalence, 15%). Mycobacteria were detected in 24 samples (86%) using the MGIT and in 23 samples (82%) using RGM medium (P = 1.00). In this setting, RGM medium showed sensitivity equivalent to that of the MGIT for isolation of NTM from the sputum of patients with CF. RGM medium offers a simple, convenient tool that can be embedded into routine culture methods, allowing the culture of all sputum samples that are submitted from patients with CF. PMID:27225412

  20. Development of sample clean up methods for the analysis of Mycobacterium tuberculosis methyl mycocerosate biomarkers in sputum extracts by gas chromatography–mass spectrometry

    PubMed Central

    Nicoara, Simona C.; Turner, Nicholas W.; Minnikin, David E.; Lee, Oona Y.-C.; O'Sullivan, Denise M.; McNerney, Ruth; Mutetwa, Reggie; Corbett, Liz E.; Morgan, Geraint H.

    2015-01-01

    A proof of principle gas chromatography–mass spectrometry method is presented, in combination with clean up assays, aiming to improve the analysis of methyl mycocerosate tuberculosis biomarkers from sputum. Methyl mycocerosates are generated from the transesterification of phthiocerol dimycocerosates (PDIMs), extracted in petroleum ether from sputum of tuberculosis suspect patients. When a high matrix background is present in the sputum extracts, the identification of the chromatographic peaks corresponding to the methyl derivatives of PDIMs analytes may be hindered by the closely eluting methyl ether of cholesterol, usually an abundant matrix constituent frequently present in sputum samples. The purification procedures involving solid phase extraction (SPE) based methods with both commercial Isolute-Florisil cartridges, and purpose designed molecularly imprinted polymeric materials (MIPs), resulted in cleaner chromatograms, while the mycocerosates are still present. The clean-up performed on solutions of PDIMs and cholesterol standards in petroleum ether show that, depending on the solvent mix and on the type of SPE used, the recovery of PDIMs is between 64 and 70%, whilst most of the cholesterol is removed from the system. When applied to petroleum ether extracts from representative sputum samples, the clean-up procedures resulted in recoveries of 36–68% for PDIMs, allowing some superior detection of the target analytes. PMID:25728371

  1. Clinical Correlates and Drug Resistance in HIV-Infected and -Uninfected Pulmonary Tuberculosis Patients in South India

    PubMed Central

    Sara, Chandy; Elsa, Heylen; Baijayanti, Mishra; Lennartsdotter, Ekstrand Maria

    2016-01-01

    Objectives To examine demographics, clinical correlates, sputum AFB (acid fast bacilli) smear grading DOTS (Directly Observed Therapy Short Course) uptake, and drug resistance in a cohort of newly-diagnosed, smear positive pulmonary tuberculosis (TB) patients with respect to HIV status at baseline, and compare smear conversion rates, side effects and mortality after two months. Design A prospective study among 54 HIV positive and 41 HIV negative pulmonary TB patients. Data were collected via face-to-face interviews, review of medical records, and lab tests. Results HIVTB co-infected patients, though more symptomatic at baseline, showed more improvement in their symptoms compared to HIV-uninfected TB patients at follow-up. The HIV co-infected group had more prevalent perceived side effects, and sputum smear positivity was marginally higher compared to the HIV negative group at follow-up. Mortality was higher among the HIV-infected group. Both groups had high rates of resistance to first-line anti-tubercular drugs, particularly isoniazid. There was no significant difference in the drug resistance patterns between the groups. Conclusions Prompt initiation and provision of daily regimens of ATT (Anti-Tubercular treatment) along with ART (Anti-Retroviral treatment) via ART centers is urgently needed in India. As resistance to ART and/or ATT is directly linked to medication non-adherence, the use of counseling, regular reinforcement, early detection and appropriate intervention strategies to tackle this complex issue could help prevent premature mortality and development of resistance in HIV-TB co-infected patients. The high rate of isoniazid resistance might preclude its use in India as prophylaxis for latent TB in HIV infected persons as per the World Health Organization (WHO) guideline.

  2. Clinical Correlates and Drug Resistance in HIV-Infected and -Uninfected Pulmonary Tuberculosis Patients in South India

    PubMed Central

    Sara, Chandy; Elsa, Heylen; Baijayanti, Mishra; Lennartsdotter, Ekstrand Maria

    2016-01-01

    Objectives To examine demographics, clinical correlates, sputum AFB (acid fast bacilli) smear grading DOTS (Directly Observed Therapy Short Course) uptake, and drug resistance in a cohort of newly-diagnosed, smear positive pulmonary tuberculosis (TB) patients with respect to HIV status at baseline, and compare smear conversion rates, side effects and mortality after two months. Design A prospective study among 54 HIV positive and 41 HIV negative pulmonary TB patients. Data were collected via face-to-face interviews, review of medical records, and lab tests. Results HIVTB co-infected patients, though more symptomatic at baseline, showed more improvement in their symptoms compared to HIV-uninfected TB patients at follow-up. The HIV co-infected group had more prevalent perceived side effects, and sputum smear positivity was marginally higher compared to the HIV negative group at follow-up. Mortality was higher among the HIV-infected group. Both groups had high rates of resistance to first-line anti-tubercular drugs, particularly isoniazid. There was no significant difference in the drug resistance patterns between the groups. Conclusions Prompt initiation and provision of daily regimens of ATT (Anti-Tubercular treatment) along with ART (Anti-Retroviral treatment) via ART centers is urgently needed in India. As resistance to ART and/or ATT is directly linked to medication non-adherence, the use of counseling, regular reinforcement, early detection and appropriate intervention strategies to tackle this complex issue could help prevent premature mortality and development of resistance in HIV-TB co-infected patients. The high rate of isoniazid resistance might preclude its use in India as prophylaxis for latent TB in HIV infected persons as per the World Health Organization (WHO) guideline. PMID:27708985

  3. Direct detection of Mycobacterium tuberculosis in sputum: A validation study using solid phase extraction-gas chromatography-mass spectrometry.

    PubMed

    Mourão, Marta P B; Kuijper, Sjoukje; Dang, Ngoc A; Walters, Elisabetta; Janssen, Hans-Gerd; Kolk, Arend H J

    2016-02-15

    Tuberculosis (TB) remains a worldwide health problem, especially in developing countries. Correct identification of Mycobacterium tuberculosis (MTB) infection is extremely important for providing appropriate treatment and care to patients. Here we describe a solid phase extraction-gas chromatography-mass spectrometry method (SPE-THM-GC-MS) for the detection of five biomarkers for M. tuberculosis. The method for classification is developed and validated through the analysis of 112 sputum samples from patients suspected of having TB. Twenty of twenty-five MTB culture-positive sputum samples were correctly classified as positive by our improved SPE-THM-GC-MS method. Eighty-five of eighty-seven MTB culture-negative samples were also negative by SPE-THM-GC-MS. The overall sensitivity of the new SPE-THM-GC-MS method is 80% (20/25) and the specificity is 98% (85/87) compared with culture. The method proved to be reliable and, although complex in principle, easy to operate due to the high degree of automation. PMID:26807702

  4. Allele-specific rpoB PCR assays for detection of rifampin-resistant Mycobacterium tuberculosis in sputum smears.

    PubMed

    Mokrousov, Igor; Otten, Tatiana; Vyshnevskiy, Boris; Narvskaya, Olga

    2003-07-01

    We describe an allele-specific PCR assay to detect mutations in three codons of the rpoB gene (516, 526, and 531) in Mycobacterium tuberculosis strains; mutations in these codons are reported to account for majority of M. tuberculosis clinical isolates resistant to rifampin (RIF), a marker of multidrug-resistant tuberculosis (MDR-TB). Three different allele-specific PCRs are carried out either directly with purified DNA (single-step multiplex allele-specific PCR), or with preamplified rpoB fragment (nested allele-specific PCR [NAS-PCR]). The method was optimized and validated following analysis of 36 strains with known rpoB sequence. A retrospective analysis of the 287 DNA preparations from epidemiologically unlinked RIF-resistant clinical strains from Russia, collected from 1996 to 2002, revealed that 247 (86.1%) of them harbored a mutation in one of the targeted rpoB codons. A prospective study of microscopy-positive consecutive sputum samples from new and chronic TB patients validated the method for direct analysis of DNA extracted from sputum smears. The potential of the NAS-PCR to control for false-negative results due to lack of amplification was proven especially useful in the study of these samples. The developed rpoB-PCR assay can be used in clinical laboratories to detect RIF-resistant and hence MDR M. tuberculosis in the regions with high burdens of the MDR-TB. PMID:12821473

  5. Serial image analysis of Mycobacterium tuberculosis colony growth reveals a persistent subpopulation in sputum during treatment of pulmonary TB.

    PubMed

    Barr, David A; Kamdolozi, Mercy; Nishihara, Yo; Ndhlovu, Victor; Khonga, Margaret; Davies, Geraint R; Sloan, Derek J

    2016-05-01

    Faster elimination of drug tolerant 'persister' bacteria may shorten treatment of tuberculosis (TB) but no method exists to quantify persisters in clinical samples. We used automated image analysis to assess whether studying growth characteristics of individual Mycobacterium tuberculosis colonies from sputum on solid media during early TB treatment facilitates 'persister' phenotyping. As Time to Detection (TTD) in liquid culture inversely correlates with total bacterial load we also evaluated the relationship between individual colony growth parameters and TTD. Sputum from TB patients in Malawi was prepared for solid and liquid culture after 0, 2 and 4 weeks of treatment. Serial photography of agar plates was used to measure time to appearance (lag time) and radial growth rate for each colony. Mixed-effects modelling was used to analyse changing growth characteristics from serial samples. 20 patients had colony measurements recorded at ≥1 time-point. Overall lag time increased by 6.5 days between baseline and two weeks (p = 0.0001). Total colony count/ml showed typical biphasic elimination, but long lag time colonies (>20days) had slower, monophasic decline. TTD was associated with minimum lag time (time to appearance of first colony1). Slower elimination of long lag time colonies suggests that these may represent a persister subpopulation of bacilli.

  6. Direct detection of Mycobacterium tuberculosis in sputum: A validation study using solid phase extraction-gas chromatography-mass spectrometry.

    PubMed

    Mourão, Marta P B; Kuijper, Sjoukje; Dang, Ngoc A; Walters, Elisabetta; Janssen, Hans-Gerd; Kolk, Arend H J

    2016-02-15

    Tuberculosis (TB) remains a worldwide health problem, especially in developing countries. Correct identification of Mycobacterium tuberculosis (MTB) infection is extremely important for providing appropriate treatment and care to patients. Here we describe a solid phase extraction-gas chromatography-mass spectrometry method (SPE-THM-GC-MS) for the detection of five biomarkers for M. tuberculosis. The method for classification is developed and validated through the analysis of 112 sputum samples from patients suspected of having TB. Twenty of twenty-five MTB culture-positive sputum samples were correctly classified as positive by our improved SPE-THM-GC-MS method. Eighty-five of eighty-seven MTB culture-negative samples were also negative by SPE-THM-GC-MS. The overall sensitivity of the new SPE-THM-GC-MS method is 80% (20/25) and the specificity is 98% (85/87) compared with culture. The method proved to be reliable and, although complex in principle, easy to operate due to the high degree of automation.

  7. Serial image analysis of Mycobacterium tuberculosis colony growth reveals a persistent subpopulation in sputum during treatment of pulmonary TB

    PubMed Central

    Barr, David A.; Kamdolozi, Mercy; Nishihara, Yo; Ndhlovu, Victor; Khonga, Margaret; Davies, Geraint R.; Sloan, Derek J.

    2016-01-01

    Summary Faster elimination of drug tolerant ‘persister’ bacteria may shorten treatment of tuberculosis (TB) but no method exists to quantify persisters in clinical samples. We used automated image analysis to assess whether studying growth characteristics of individual Mycobacterium tuberculosis colonies from sputum on solid media during early TB treatment facilitates ‘persister’ phenotyping. As Time to Detection (TTD) in liquid culture inversely correlates with total bacterial load we also evaluated the relationship between individual colony growth parameters and TTD. Sputum from TB patients in Malawi was prepared for solid and liquid culture after 0, 2 and 4 weeks of treatment. Serial photography of agar plates was used to measure time to appearance (lag time) and radial growth rate for each colony. Mixed-effects modelling was used to analyse changing growth characteristics from serial samples. 20 patients had colony measurements recorded at ≥1 time-point. Overall lag time increased by 6.5 days between baseline and two weeks (p = 0.0001). Total colony count/ml showed typical biphasic elimination, but long lag time colonies (>20days) had slower, monophasic decline. TTD was associated with minimum lag time (time to appearance of first colony1). Slower elimination of long lag time colonies suggests that these may represent a persister subpopulation of bacilli. PMID:27156626

  8. Induced Sputum Nitrite Levels Correlate with Clinical Asthma Parameters in Children Aged 7–18 Years with Mild to Moderate Persistent Asthma

    PubMed Central

    Nandan, Devki; Kansal, Prachi; Patharia, Neha; Goyal, Parul

    2016-01-01

    Purpose: The objective of this study is to measure levels of nitrites in induced sputum in children with asthma and correlate it with clinical asthma parameters. Method: This prospective observational study was done in PGIMER, Dr. Ram Manohar Lohia Hospital, New Delhi, on 91 children aged 7-18 years with mild and moderate persistent asthma. Patients were specifically evaluated for five clinical parameters of asthma (i.e. Days of acute exacerbations, use of salbutamol as rescue medication, emergency visits, nights with cough, days of school absence) and induced sputum nitrite levels was done at the time of enrollment and 3 months after treatment with inhaled budesonide. Results: The mean age of subjects was 10.79 ± 2.563yrs. Six (6.59%) patients were not able to perform induced sputum, eighty five (93.41%) patients were suitable for data analysis. There was significant reduction in sputum nitrite levels from 33.42 ± 22.04nmol/ml at enrollment to 11.72 ± 5.61 nmol/ml (P < 0.0005) after 3 months of inhaled budesonide therapy. Significant positive correlation was found between reduction in sputum nitrite level and control of asthma symptoms: Days of acute exacerbations(r value = 0.548, P value = 0.0001), Days of salbutamol use as rescue medication (r value = 0.431, P value =< 0.0001), Number of emergency visits(r value = 0.414, P value = 0.0001), Nights with cough (r value = 0.259, P value = 0.0169), Days of school absence(r value = 0.411, P value = 0.0001). Sputum nitrite levels were significantly higher in moderate persistent asthmatics as compared to mild at the time of enrollment (P < 0.0005), which shows that induced sputum nitrite levels correlate with asthma severity. Conclusions: This study confirms that nitrites in induced sputum correlate well with clinical asthma parameters and asthma severity in children and is a simple, non invasive, and cheap method which can be used as a parameter for monitoring of asthma. PMID:27365917

  9. [Induced sputum supernatant prostaglandin E2 during oral aspirin challenge of asthmatic patients with and without aspirin hypersensitivity and healthy controls--pilot study].

    PubMed

    Ignacak, Maria; Celejewska-Wójcik, Natalia; Wójcik, Krzysztof; Sałapa, Kinga; Konduracka, Ewa; Sanak, Marek; Tyrak, Katarzyna; Sładek, Krzysztof; Musiał, Jacek; Mastalerz, Lucyna

    2016-01-01

    The aim of this pilot study was to evaluate changes in the concentration of prostaglandin E2 (PGE2) in induced sputum supernatant in 3 groups: sub- jects with NSAID-exacerbated respira- tory disease (NERD), aspirin tolerant asthma (ATA) and healthy controls (HC), before and after oral aspirin chal- lenge test. The study was conducted in the years 2014-2015 at the Clinical Department of the Pulmonology Clinic at the University Hospital in Cracow. 43 patients were enrolled in the study (NERD - n = 15, ATA - n = 15 and HC - n = 13). All of them underwent a placebo-controlled oral aspirin challenge. Sputum was induced 24 hours before the challenge and immediately after the test. Induced sputum was processed in order to obtain cystospin slides to depict inflammatory cell patterns and supernatants, in which PGE2 was measured. The concentration of PGE2 was determined using mass spectrometry coupled with gas chromatography (gas chromatography/mass spectrometry - GC/MS). After aspirin challenge, the concentration of PGE2 in induced sputum supernatant decreased in both asthmatics hypersensitive to aspirin (p = 0.01) and those who tolerated aspirin well (p = 0.17). The change in the healthy control group was not statistically significant. These results support the cyclooxygenase theory of PGE2 inhibition by aspirin. However, the mechanism of bronchoconstriction after aspirin administration alone in patients with NSAID-exacerbated respiratory disease remains unclear.

  10. DETECTION OF K-RAS AND P53 MUTATIONS IN SPUTUM SAMPLES OF LUNG CANCER PATIENTS USING LASER CAPTURE MICRODISSECTION MICROSCOPE AND MUTATION ANALYSIS

    EPA Science Inventory

    Detection of K-ras and p53 Mutations in Sputum Samples of Lung Cancer Patients Using Laser Capture Microdissection Microscope and Mutation Analysis

    Phouthone Keohavong a,*, Wei-Min Gao a, Kui-Cheng Zheng a, Hussam Mady b, Qing Lan c, Mona Melhem b, and Judy Mumford d.
    <...

  11. [Induced sputum supernatant prostaglandin E2 during oral aspirin challenge of asthmatic patients with and without aspirin hypersensitivity and healthy controls--pilot study].

    PubMed

    Ignacak, Maria; Celejewska-Wójcik, Natalia; Wójcik, Krzysztof; Sałapa, Kinga; Konduracka, Ewa; Sanak, Marek; Tyrak, Katarzyna; Sładek, Krzysztof; Musiał, Jacek; Mastalerz, Lucyna

    2016-01-01

    The aim of this pilot study was to evaluate changes in the concentration of prostaglandin E2 (PGE2) in induced sputum supernatant in 3 groups: sub- jects with NSAID-exacerbated respira- tory disease (NERD), aspirin tolerant asthma (ATA) and healthy controls (HC), before and after oral aspirin chal- lenge test. The study was conducted in the years 2014-2015 at the Clinical Department of the Pulmonology Clinic at the University Hospital in Cracow. 43 patients were enrolled in the study (NERD - n = 15, ATA - n = 15 and HC - n = 13). All of them underwent a placebo-controlled oral aspirin challenge. Sputum was induced 24 hours before the challenge and immediately after the test. Induced sputum was processed in order to obtain cystospin slides to depict inflammatory cell patterns and supernatants, in which PGE2 was measured. The concentration of PGE2 was determined using mass spectrometry coupled with gas chromatography (gas chromatography/mass spectrometry - GC/MS). After aspirin challenge, the concentration of PGE2 in induced sputum supernatant decreased in both asthmatics hypersensitive to aspirin (p = 0.01) and those who tolerated aspirin well (p = 0.17). The change in the healthy control group was not statistically significant. These results support the cyclooxygenase theory of PGE2 inhibition by aspirin. However, the mechanism of bronchoconstriction after aspirin administration alone in patients with NSAID-exacerbated respiratory disease remains unclear. PMID:27197430

  12. A fast method for the identification of Mycobacterium tuberculosis in sputum and cultures based on thermally assisted hydrolysis and methylation followed by gas chromatography-mass spectrometry.

    PubMed

    Kaal, Erwin; Kolk, Arend H J; Kuijper, Sjoukje; Janssen, Hans-Gerd

    2009-08-28

    A fast gas chromatography-mass spectrometry (GC-MS) method with minimum sample preparation is described for early diagnosis of tuberculosis (TB). The automated procedure is based on the injection of sputum samples which are then methylated inside the GC injector using thermally assisted hydrolysis and methylation (THM). The THM-GC-MS procedure was optimized for the injection of sputum samples. For the identification of Mycobacterium tuberculosis the known marker tuberculostearic acid (TBSA) and other potential markers were evaluated. Hexacosanoic acid in combination with TBSA was found to be specific for the presence of M. tuberculosis. For validation of the method several sputum samples with different viscosities spiked with bacterial cultures were analyzed. Finally, 18 stored sputum samples collected in Vietnam from patients suspected to suffer from TB were re-analyzed in Amsterdam by microscopy after decontamination/concentration and using the new THM-GC-MS method. No false positives were found by THM-GC-MS and all patients who were diagnosed with TB were also found positive using our newly developed THM-GC-MS method. These results show that the new fast and sensitive THM-GC-MS method holds great potential for the diagnosis of TB.

  13. DOUBLE-STEP MULTIPLEX REAL TIME PCR WITH MELTING CURVE ANALYSIS FOR DETECTION AND DIFFERENTIATION OF MYCOBACTERIA IN SPUTUM.

    PubMed

    Kasa, Sawinee; Faksri, Kiatichai; Kaewkes, Wanlop; Lulitanond, Viraphong; Namwat, Wises

    2015-01-01

    Mycobacterium tuberculosis (M. tb) is a causative agent of tuberculosis, a worldwide public health problem. In recent years, the incidence of human mycobacterial infection due to species other than M. tb has increased. However, the lack of specific, rapid, and inexpensive methods for identification of mycobacterial species remains a pressing problem. A diagnostic test was developed for mycobacterial strain differentiation utilizing a double-step multiplex real time PCR together with melting curve analysis for identifying and distinguishing among M. tb, M. bovis BCG, other members of M. tb. complex, M. avium, and non-tuberculosis mycobacteria. The assay was tested using 167 clinical sputum samples in comparison with acid-fast staining and culturing. Using only the first step (step A) the assay achieved sensitivity and specificity of 81% and 95%, respectively. The detection limit was equivalent to 50 genome copies.

  14. DOUBLE-STEP MULTIPLEX REAL TIME PCR WITH MELTING CURVE ANALYSIS FOR DETECTION AND DIFFERENTIATION OF MYCOBACTERIA IN SPUTUM.

    PubMed

    Kasa, Sawinee; Faksri, Kiatichai; Kaewkes, Wanlop; Lulitanond, Viraphong; Namwat, Wises

    2015-01-01

    Mycobacterium tuberculosis (M. tb) is a causative agent of tuberculosis, a worldwide public health problem. In recent years, the incidence of human mycobacterial infection due to species other than M. tb has increased. However, the lack of specific, rapid, and inexpensive methods for identification of mycobacterial species remains a pressing problem. A diagnostic test was developed for mycobacterial strain differentiation utilizing a double-step multiplex real time PCR together with melting curve analysis for identifying and distinguishing among M. tb, M. bovis BCG, other members of M. tb. complex, M. avium, and non-tuberculosis mycobacteria. The assay was tested using 167 clinical sputum samples in comparison with acid-fast staining and culturing. Using only the first step (step A) the assay achieved sensitivity and specificity of 81% and 95%, respectively. The detection limit was equivalent to 50 genome copies. PMID:26513906

  15. A Review of Automatic Methods Based on Image Processing Techniques for Tuberculosis Detection from Microscopic Sputum Smear Images.

    PubMed

    Panicker, Rani Oomman; Soman, Biju; Saini, Gagan; Rajan, Jeny

    2016-01-01

    Tuberculosis (TB) is an infectious disease caused by the bacteria Mycobacterium tuberculosis. It primarily affects the lungs, but it can also affect other parts of the body. TB remains one of the leading causes of death in developing countries, and its recent resurgences in both developed and developing countries warrant global attention. The number of deaths due to TB is very high (as per the WHO report, 1.5 million died in 2013), although most are preventable if diagnosed early and treated. There are many tools for TB detection, but the most widely used one is sputum smear microscopy. It is done manually and is often time consuming; a laboratory technician is expected to spend at least 15 min per slide, limiting the number of slides that can be screened. Many countries, including India, have a dearth of properly trained technicians, and they often fail to detect TB cases due to the stress of a heavy workload. Automatic methods are generally considered as a solution to this problem. Attempts have been made to develop automatic approaches to identify TB bacteria from microscopic sputum smear images. In this paper, we provide a review of automatic methods based on image processing techniques published between 1998 and 2014. The review shows that the accuracy of algorithms for the automatic detection of TB increased significantly over the years and gladly acknowledges that commercial products based on published works also started appearing in the market. This review could be useful to researchers and practitioners working in the field of TB automation, providing a comprehensive and accessible overview of methods of this field of research. PMID:26573654

  16. Synthetic Cystic Fibrosis Sputum Medium Regulates Flagellar Biosynthesis through the flhF Gene in Burkholderia cenocepacia.

    PubMed

    Kumar, Brijesh; Cardona, Silvia T

    2016-01-01

    Burkholderia cenocepacia belongs to the Burkholderia cepacia complex (Bcc), a group of at least 18 distinct species that establish chronic infections in the lung of people with the genetic disease cystic fibrosis (CF). The sputum of CF patients is rich in amino acids and was previously shown to increase flagellar gene expression in B. cenocepacia. We examined flagellin expression and flagellar morphology of B. cenocepacia grown in synthetic cystic fibrosis sputum medium (SCFM) compared to minimal medium. We found that CF nutritional conditions induce increased motility and flagellin expression. Individual amino acids added at the same concentrations as found in SCFM also increased motility but not flagellin expression, suggesting a chemotactic effect of amino acids. Electron microscopy and flagella staining demonstrated that the increase in flagellin corresponds to a change in the number of flagella per cell. In minimal medium, the ratio of multiple: single: aflagellated cells was 2:3.5:4.5; while under SCFM conditions, the ratio was 7:2:1. We created a deletion mutant, ΔflhF, to study whether this putative GTPase regulates the flagellation pattern of B. cenocepacia K56-2 during growth in CF conditions. The ΔflhF mutant exhibited 80% aflagellated, 14% single and 6% multiple flagellated bacterial subpopulations. Moreover, the ratio of multiple to single flagella in WT and ΔflhF was 3.5 and 0.43, respectively in CF conditions. The observed differences suggest that FlhF positively regulates flagellin expression and the flagellation pattern in B. cenocepacia K56-2 during CF nutritional conditions. PMID:27379216

  17. Synthetic Cystic Fibrosis Sputum Medium Regulates Flagellar Biosynthesis through the flhF Gene in Burkholderia cenocepacia

    PubMed Central

    Kumar, Brijesh; Cardona, Silvia T.

    2016-01-01

    Burkholderia cenocepacia belongs to the Burkholderia cepacia complex (Bcc), a group of at least 18 distinct species that establish chronic infections in the lung of people with the genetic disease cystic fibrosis (CF). The sputum of CF patients is rich in amino acids and was previously shown to increase flagellar gene expression in B. cenocepacia. We examined flagellin expression and flagellar morphology of B. cenocepacia grown in synthetic cystic fibrosis sputum medium (SCFM) compared to minimal medium. We found that CF nutritional conditions induce increased motility and flagellin expression. Individual amino acids added at the same concentrations as found in SCFM also increased motility but not flagellin expression, suggesting a chemotactic effect of amino acids. Electron microscopy and flagella staining demonstrated that the increase in flagellin corresponds to a change in the number of flagella per cell. In minimal medium, the ratio of multiple: single: aflagellated cells was 2:3.5:4.5; while under SCFM conditions, the ratio was 7:2:1. We created a deletion mutant, ΔflhF, to study whether this putative GTPase regulates the flagellation pattern of B. cenocepacia K56-2 during growth in CF conditions. The ΔflhF mutant exhibited 80% aflagellated, 14% single and 6% multiple flagellated bacterial subpopulations. Moreover, the ratio of multiple to single flagella in WT and ΔflhF was 3.5 and 0.43, respectively in CF conditions. The observed differences suggest that FlhF positively regulates flagellin expression and the flagellation pattern in B. cenocepacia K56-2 during CF nutritional conditions. PMID:27379216

  18. MicroRNA expression profiling of sputum for the detection of early and locally advanced non-small-cell lung cancer: a prospective case–control study

    PubMed Central

    Razzak, R.; Bédard, E.L.R.; Kim, J.O.; Gazala, S.; Guo, L.; Ghosh, S.; Joy, A.; Nijjar, T.; Wong, E.; Roa, W.H.

    2016-01-01

    Background Non-small-cell lung cancer (nsclc) is associated with very poor overall survival because 70% of patients present with locally advanced or metastatic disease at the time of diagnosis. Micrornas (mirnas) are a class of short, noncoding rna molecules whose presence in samples of biologic fluids such as sputum has demonstrated promise as a potential means of detecting nsclc. We investigated the stage-specific nsclc detection potential of an efficient panel of 3 mirnas (mir-21, mir-210, mir-372) using a single sputum sample. Methods A single spontaneously expectorated sputum sample was prospectively collected from 21 early nsclc (≤stage ii) patients, 22 advanced nsclc (≥stage iii) patients, and 10 control subjects. Mirna expression profiles were determined by quantitative real-time polymerase chain reaction and were analyzed by unsupervised hierarchical cluster analysis. Results Mean tumour size (±95% confidence interval) in the early and advanced nsclc patients was 3.3 cm ± 0.9 cm and 4.8 cm ± 0.7 cm respectively. Adenocarcinoma constituted 61.9% of the early and 45.5% of the advanced nsclc cases respectively. In comparing the early nsclc group with the control group, the mirna panel yielded a diagnostic sensitivity of 67% and a specificity of 90.0%. For the advanced nsclc group, the mirna panel detected nsclc with a sensitivity and specificity of 64% and 100% respectively. Conclusions A sputum mir-21, mir-210, and mir-372 expression profile might provide a sensitive and highly specific means for detecting nsclc. Sputum mirna analysis demonstrates promise as a potential complementary screening tool. PMID:27122989

  19. First Airswot Interferometric Radar Water Surface Elevations and Flooded Inundation Extent from the Sacramento River and Edwards AFB Wetland Complex, California

    NASA Astrophysics Data System (ADS)

    Pitcher, L. H.; Smith, L. C.; Gleason, C. J.; Baney, O. N.; Chu, V. W.; Bennett, M. M.; Pavelsky, T.; Sadowy, G. A.

    2014-12-01

    NASA's forthcoming Surface Water Ocean Topography (SWOT) satellite mission aims to quantify global freshwater fluxes from space using Ka-band interferometric radar. AirSWOT is the airborne calibration/validation instrument for SWOT with first-pass data collected over the Sacramento River in May 2013 and a wetland complex on Edwards AFB (Piute Ponds) in May 2014. Here, AirSWOT elevation and coherence data are compared with high resolution airborne imagery and concurrent in-situ field mappings of inundation area and water surface elevation. For the Sacramento River, AirSWOT water surface elevations are compared with field-surveyed elevations collected using a high precision GPS Lagrangian river drifter escorted down 30 km of river length. Additionally, field mapped river shorelines are compared with shorelines extracted from AirSWOT coherence data. For the Piute Ponds, we use an exhaustive field mapping of inundation extent and flooded vegetation to assess the ability of AirSWOT coherence and backscatter to map shorelines in a complex lake and wetland environment containing varying vegetation and soil moisture conditions.

  20. Measurement of the Inclusive Forward-Backward top quark-antitop quark Production Asymmetry and its Rapidity Dependence dAfb/d(delta y)

    NASA Astrophysics Data System (ADS)

    Strycker, Glenn Lloyd

    Early measurements of a large forward-background asymmetry at the CDF [1, 2, 3, 4] and Dempty [5, 6] experiments at Fermilab have generated much recent interest, but were hampered by large uncertainties. We present here a new measurement of the corrected forward-backward asymmetry of pair-produced top quarks, using a high-statistics sample with much improved precision. We study the rapidity, ytop, of the top quark production angle with respect to the incoming parton momentum in both the lab and tt¯ rest frames. We find the corrected forward-backward asymmetries to be Appfb=0.150+/- 0.050stat+/-0.024syst Attfb=0.158+/-0 .072stat+/-0.024syst These results should be compared with the small lab pp¯ frame charge asymmetry expected in QCD at NLO, Afb = 0.050 +/- 0.015 [7, 8, 9, 10]. Additionally, we introduce a measurement of the A fb rapidity dependence dAfbd Dy . We find this to be Attfb Dy<1.0 =0.026+/-0.104stat+/-0.012 syst Attfb Dy>1.0=0.6 11+/-0.210stat+/-0.246syst which we compare with model predictions 0.039 +/- 0.006 and 0.123 +/- 0.018 for the inner and outer rapidities, respectively.

  1. Superfund Record of Decision (EPA Region 10): American Lake Gardens (McCord AFB - Area D), Pierce County, WA. (First remedial action), September 1991. Final report

    SciTech Connect

    Not Available

    1991-09-19

    The American Lake Gardens (McChord AFB-Area D) site is an active U.S. Air Force base located at McChord Air Force Base, Pierce County, Washington. The site consists of two areas, Area D and American Lake Garden Tract (ALGT). From the mid-1940's to the present, no known industrial activities have occurred in the ALGT area; however, seven waste disposal sites have operated within the Area D portion of the site. Concurrent with DOD investigations, EPA discovered TCE in ground water monitoring wells installed at the ALGT, and in 1984, concluded that waste disposal sites in Area D were the likely source of ground water contamination. The ROD addresses remediation of the contaminated onsite and offsite ground water plume, as a final remedy. The primary contaminants of concern affecting the ground water are VOCs including benzene, PCE, TCE, toluene, and xylenes; other organics; and metals including arsenic, chromium, and lead. The selected remedial action for the site is included.

  2. The E3 Ubiquitin Ligase SCFTIR1/AFB and Membrane Sterols Play Key Roles in Auxin Regulation of Endocytosis, Recycling, and Plasma Membrane Accumulation of the Auxin Efflux Transporter PIN2 in Arabidopsis thaliana[C][W][OA

    PubMed Central

    Pan, Jianwei; Fujioka, Shozo; Peng, Jianling; Chen, Jianghua; Li, Guangming; Chen, Rujin

    2009-01-01

    The PIN family of auxin efflux transporters exhibit polar plasma membrane (PM) localization and play a key role in auxin gradient-mediated developmental processes. Auxin inhibits PIN2 endocytosis and promotes its PM localization. However, the underlying mechanisms remain elusive. Here, we show that the inhibitory effect of auxin on PIN2 endocytosis was impaired in SCFTIR1/AFB auxin signaling mutants. Similarly, reducing membrane sterols impaired auxin inhibition of PIN2 endocytosis. Gas chromatography–mass spectrometry analyses indicate that membrane sterols were significantly reduced in SCFTIR1/AFB mutants, supporting a link between membrane sterols and auxin signaling in regulating PIN2 endocytosis. We show that auxin promoted PIN2 recycling from endosomes to the PM and increased PIN2 steady state levels in the PM fraction. Furthermore, we show that the positive effect of auxin on PIN2 levels in the PM was impaired by inhibiting membrane sterols or auxin signaling. Consistent with this, the sterol biosynthetic mutant fk-J79 exhibited pronounced defects in primary root elongation and gravitropic response. Our data collectively indicate that, although there are distinct processes involved in endocytic regulation of specific PM-resident proteins, the SCFTIR1/AFB-dependent processes are required for auxin regulation of endocytosis, recycling, and PM accumulation of the auxin efflux transporter PIN2 in Arabidopsis thaliana. PMID:19218398

  3. The effect of forced expiration on the uniformity of 99Tcm-DTPA aerosol ventilation images in patients with excess sputum production.

    PubMed

    Middleton, E; Clout, C; Occleshaw, C; Tindale, W B; Parker, B V; Barber, D C; Barrington, N A

    1990-08-01

    This study examines the use of the forced expiratory technique (FET) as a means of improving the uniformity of radionuclide aerosol ventilation images in patients with excess sputum production. Ventilation images are objectively classified by two computer derived indices to characterize the degree of radioaerosol clumping and overall uniformity. In a series of twenty five patients with a long-standing history of daily sputum production, images acquired before and after forced expiration and again after a second ventilation immediately following FET showed no significant change in either index. The results obtained do not provide evidence to support the routine use of FET in conjunction with radioaerosol scintigraphy. The method of image classification correlates well with a visual assessment of image uniformity and has general application.

  4. Detection and Quantification of Mycobacterium tuberculosis in the Sputum of Culture-Negative HIV-infected Pulmonary Tuberculosis Suspects: A Proof-of-Concept Study

    PubMed Central

    Madico, Guillermo; Mpeirwe, Moses; White, Laura; Vinhas, Solange; Orr, Beverley; Orikiriza, Patrick; Miller, Nancy S.; Gaeddert, Mary; Mwanga-Amumpaire, Juliet; Palaci, Moises; Kreiswirth, Barry; Straight, Joe; Dietze, Reynaldo; Boum, Yap; Jones-López, Edward C.

    2016-01-01

    Rationale Rapid diagnosis of pulmonary tuberculosis (TB) is critical for timely initiation of treatment and interruption of transmission. Yet, despite recent advances, many patients remain undiagnosed. Culture, usually considered the most sensitive diagnostic method, is sub-optimal for paucibacillary disease. Methods We evaluated the Totally Optimized PCR (TOP) TB assay, a new molecular test that we hypothesize is more sensitive than culture. After pre-clinical studies, we estimated TOP’s per-patient sensitivity and specificity in a convenience sample of 261 HIV-infected pulmonary TB suspects enrolled into a TB diagnostic study in Mbarara, Uganda against MGIT culture, Xpert MTB/RIF and a composite reference standard. We validated results with a confirmatory PCR used for sequencing M. tuberculosis. Measurements and Results Using culture as reference, TOP had 100% sensitivity but 35% specificity. Against a composite reference standard, the sensitivity of culture (27%) and Xpert MTB/RIF (27%) was lower than TOP (99%), with similar specificity (100%, 98% and 87%, respectively). In unadjusted analyses, culture-negative/TOP-positive patients were more likely to be older (P<0·001), female (P<0·001), have salivary sputum (P = 0·05), sputum smear-negative (P<0.001) and less advanced disease on chest radiograph (P = 0.05). M. tuberculosis genotypes identified in sputum by DNA sequencing exhibit differential growth in culture. Conclusions These findings suggest that the TOP TB assay is accurately detecting M. tuberculosis DNA in the sputum of culture-negative tuberculosis suspects. Our results require prospective validation with clinical outcomes. If the operating characteristics of the TOP assay are confirmed in future studies, it will be justified as a “TB rule out” test. PMID:27391604

  5. Responsiveness of blood and sputum inflammatory cells in Japanese COPD patients, non-COPD smoking controls, and non-COPD nonsmoking controls

    PubMed Central

    Kawayama, Tomotaka; Kinoshita, Takashi; Matsunaga, Kazuko; Kobayashi, Akihiro; Hayamizu, Tomoyuki; Johnson, Malcolm; Hoshino, Tomoaki

    2016-01-01

    Purpose To compare pulmonary and systemic inflammatory mediator release, pre- and poststimulation, ex vivo, in cells from Japanese patients with chronic obstructive pulmonary disease (COPD), non-COPD smoking controls, and non-COPD nonsmoking controls (NSC). Patients and methods This was a nontreatment study with ten subjects per group. Inflammatory biomarker release, including interleukin (IL)-6 and -8, matrix metalloproteinase-9, and tumor necrosis factor (TNF)-α, was measured in peripheral blood mononuclear cells (PBMC) and sputum cells with and without lipopolysaccharide or TNF-α stimulation. Results In PBMC, basal TNF-α release (mean ± standard deviation) was significantly different between COPD (81.6±111.4 pg/mL) and nonsmoking controls (9.5±5.2 pg/mL) (P<0.05). No other significant differences were observed. Poststimulation biomarker release tended to increase, with the greatest changes in the COPD group. The greatest mean increases were seen in the lipopolysaccharide-induced release of matrix metalloproteinase-9, TNF-α, and IL-6 from PBMC. Pre- and poststimulation data from sputum samples were more variable and less conclusive than from PBMC. In the COPD group, induced sputum neutrophil levels were higher and macrophage levels were lower than in either control group. Significant correlations were seen between the number of sputum cells (macrophages and neutrophils) and biomarker levels (IL-8, IL-6, and TNF-α). Conclusion This was the first study to compare cellular inflammatory mediator release before and after stimulation among Japanese COPD, smoking controls, and nonsmoking controls populations. Poststimulation levels tended to be higher in patients with COPD. The results suggest that PBMC are already preactivated in the circulation in COPD patients. This provides further evidence that COPD is a multicomponent disease, involving both airway and systemic inflammation. PMID:26929615

  6. Effects of piclamilast, a selective phosphodiesterase-4 inhibitor, on oxidative burst of sputum cells from mild asthmatics and stable COPD patients.

    PubMed

    Beeh, Kai M; Beier, Jutta; Lerch, Claudia; Schulz, Ann K; Buhl, Roland

    2004-01-01

    Oxidative stress associated with increased presence of neutrophils is an important feature of inflammatory airways diseases like asthma or chronic obstructive pulmonary disease. We studied the in vitro effect of piclamilast (RP73401), a selective phosphodiesterase (PDE)-4 inhibitor, compared to theophylline and prednisolone, on respiratory burst of sputum cells from mild asthmatics and COPD patients. Sputum cells were harvested from mild asthmatics and stable COPD patients and treated with piclamilast, theophylline or prednisolone. Respiratory burst was assessed by luminol-dependent chemoluminescence after stimulation with 10 microM n-formyl-met-leu-phe (FMLP). Piclamilast inhibited FMLP-induced respiratory burst of sputum cells in a concentration-dependent manner (asthma: EC50 approximately 100 nM, max. inhibition: 97.5+/-5% at 100 microM; COPD: EC50 approximately 1 microM, max. inhibition: 70.6+/-4.5% at 100 microM), whereas maximal inhibition observed with theophylline (asthma: max. inhib. 27+/-15%; COPD: 6+/-2%, both p < 0.05 vs. piclamilast) and prednisolone (asthma: 16+/-6%; COPD: 7.8+/-6.2%, both p < 0.05 vs. piclamilast) was weaker. Inhibition by piclamilast was largely reversed through pretreatment of cells with the adenylcyclase inhibitor SQ22536. We concluded that piclamilast, a selective PDE-4 inhibitor, attenuates the respiratory burst of sputum cells from mild asthmatics and COPD patients in vitro. These data underline the potential of PDE-4 inhibition as a novel therapeutic approach to inflammatory airway diseases like asthma or COPD.

  7. A Subset of Circulating Blood Mycobacteria-Specific CD4 T Cells Can Predict the Time to Mycobacterium tuberculosis Sputum Culture Conversion

    PubMed Central

    Lugongolo, Masixole; Gwala, Thabisile; Kiravu, Agano; Deniso, Pamela; Stewart-Isherwood, Lynsey; Omar, Shaheed Vally; Grobusch, Martin P.; Coetzee, Gerrit; Conradie, Francesca; Ismail, Nazir; Kaplan, Gilla; Fallows, Dorothy

    2014-01-01

    We investigated 18 HIV-negative patients with MDR-TB for M. tuberculosis (Mtb)- and PPD-specific CD4 T cell responses and followed them over 6 months of drug therapy. Twelve of these patients were sputum culture (SC) positive and six patients were SC negative upon enrollment. Our aim was to identify a subset of mycobacteria-specific CD4 T cells that would predict time to culture conversion. The total frequency of mycobacteria-specific CD4 T cells at baseline could not distinguish patients showing positive or negative SC. However, a greater proportion of late-differentiated (LD) Mtb- and PPD-specific memory CD4 T cells was found in SC positive patients than in those who were SC negative (p = 0.004 and p = 0.0012, respectively). Similarly, a higher co-expression of HLA-DR+Ki67+ on Mtb- and PPD-specific CD4 T cells could also discriminate between sputum SC positive versus SC negative (p = 0.004 and p = 0.001, respectively). Receiver operating characteristic (ROC) analysis revealed that baseline levels of Ki67+HLA-DR+ Mtb- and PPD-specific CD4 T cells were predictive of the time to sputum culture conversion, with area-under-the-curve of 0.8 (p = 0.027). Upon treatment, there was a significant decline of these Ki67+HLA-DR+ T cell populations in the first 2 months, with a progressive increase in mycobacteria-specific polyfunctional IFNγ+IL2+TNFα+ CD4 T cells over 6 months. Thus, a subset of activated and proliferating mycobacterial-specific CD4 T cells (Ki67+HLA-DR+) may provide a valuable marker in peripheral blood that predicts time to sputum culture conversion in TB patients at the start of treatment. PMID:25048802

  8. A subset of circulating blood mycobacteria-specific CD4 T cells can predict the time to Mycobacterium tuberculosis sputum culture conversion.

    PubMed

    Riou, Catherine; Gray, Clive M; Lugongolo, Masixole; Gwala, Thabisile; Kiravu, Agano; Deniso, Pamela; Stewart-Isherwood, Lynsey; Omar, Shaheed Vally; Grobusch, Martin P; Coetzee, Gerrit; Conradie, Francesca; Ismail, Nazir; Kaplan, Gilla; Fallows, Dorothy

    2014-01-01

    We investigated 18 HIV-negative patients with MDR-TB for M. tuberculosis (Mtb)- and PPD-specific CD4 T cell responses and followed them over 6 months of drug therapy. Twelve of these patients were sputum culture (SC) positive and six patients were SC negative upon enrollment. Our aim was to identify a subset of mycobacteria-specific CD4 T cells that would predict time to culture conversion. The total frequency of mycobacteria-specific CD4 T cells at baseline could not distinguish patients showing positive or negative SC. However, a greater proportion of late-differentiated (LD) Mtb- and PPD-specific memory CD4 T cells was found in SC positive patients than in those who were SC negative (p = 0.004 and p = 0.0012, respectively). Similarly, a higher co-expression of HLA-DR+ Ki67+ on Mtb- and PPD-specific CD4 T cells could also discriminate between sputum SC positive versus SC negative (p = 0.004 and p = 0.001, respectively). Receiver operating characteristic (ROC) analysis revealed that baseline levels of Ki67+ HLA-DR+ Mtb- and PPD-specific CD4 T cells were predictive of the time to sputum culture conversion, with area-under-the-curve of 0.8 (p = 0.027). Upon treatment, there was a significant decline of these Ki67+ HLA-DR+ T cell populations in the first 2 months, with a progressive increase in mycobacteria-specific polyfunctional IFNγ+ IL2+ TNFα+ CD4 T cells over 6 months. Thus, a subset of activated and proliferating mycobacterial-specific CD4 T cells (Ki67+ HLA-DR+) may provide a valuable marker in peripheral blood that predicts time to sputum culture conversion in TB patients at the start of treatment.

  9. A subset of circulating blood mycobacteria-specific CD4 T cells can predict the time to Mycobacterium tuberculosis sputum culture conversion.

    PubMed

    Riou, Catherine; Gray, Clive M; Lugongolo, Masixole; Gwala, Thabisile; Kiravu, Agano; Deniso, Pamela; Stewart-Isherwood, Lynsey; Omar, Shaheed Vally; Grobusch, Martin P; Coetzee, Gerrit; Conradie, Francesca; Ismail, Nazir; Kaplan, Gilla; Fallows, Dorothy

    2014-01-01

    We investigated 18 HIV-negative patients with MDR-TB for M. tuberculosis (Mtb)- and PPD-specific CD4 T cell responses and followed them over 6 months of drug therapy. Twelve of these patients were sputum culture (SC) positive and six patients were SC negative upon enrollment. Our aim was to identify a subset of mycobacteria-specific CD4 T cells that would predict time to culture conversion. The total frequency of mycobacteria-specific CD4 T cells at baseline could not distinguish patients showing positive or negative SC. However, a greater proportion of late-differentiated (LD) Mtb- and PPD-specific memory CD4 T cells was found in SC positive patients than in those who were SC negative (p = 0.004 and p = 0.0012, respectively). Similarly, a higher co-expression of HLA-DR+ Ki67+ on Mtb- and PPD-specific CD4 T cells could also discriminate between sputum SC positive versus SC negative (p = 0.004 and p = 0.001, respectively). Receiver operating characteristic (ROC) analysis revealed that baseline levels of Ki67+ HLA-DR+ Mtb- and PPD-specific CD4 T cells were predictive of the time to sputum culture conversion, with area-under-the-curve of 0.8 (p = 0.027). Upon treatment, there was a significant decline of these Ki67+ HLA-DR+ T cell populations in the first 2 months, with a progressive increase in mycobacteria-specific polyfunctional IFNγ+ IL2+ TNFα+ CD4 T cells over 6 months. Thus, a subset of activated and proliferating mycobacterial-specific CD4 T cells (Ki67+ HLA-DR+) may provide a valuable marker in peripheral blood that predicts time to sputum culture conversion in TB patients at the start of treatment. PMID:25048802

  10. Potential yield of pulmonary tuberculosis cases by direct microscopy of sputum in a district of South India*

    PubMed Central

    Baily, G. V. J.; Savič, D.; Gothi, G. D.; Naidu, V. B.; Nair, S. S.

    1967-01-01

    In the District Tuberculosis Programme in India, which is part of a national programme, case-finding and treatment of pulmonary tuberculosis are integrated into the general health services. With such an integration, it is desirable, in order to forecast possible achievements, to understand the additional work-load that the programme will impose upon the existing health institutions and to assess the potential of these institutions for case-finding and treatment. The operational study described in this paper was made under programme conditions in a district in South India and is an examination of the case-finding potential of rural health institutions, using direct microscopy of sputum for diagnosis, and of the additional work-load involved by such case-finding. Under the conditions found in the course of the study, it is clear that a substantial proportion (at least 65%) of pulmonary tuberculosis cases prevalent in the district can be detected by the existing staffs with an easily manageable work-load. PMID:5301646

  11. Phenylalanine induces Burkholderia cenocepacia phenylacetic acid catabolism through degradation to phenylacetyl-CoA in synthetic cystic fibrosis sputum medium.

    PubMed

    Yudistira, Harry; McClarty, Leigh; Bloodworth, Ruhi A M; Hammond, Sydney A; Butcher, Haley; Mark, Brian L; Cardona, Silvia T

    2011-09-01

    Synthetic cystic fibrosis sputum medium (SCFM) is rich in amino acids and supports robust growth of Burkholderia cenocepacia, a member of the Burkholderia cepacia complex (Bcc). Previous work demonstrated that B. cenocepacia phenylacetic acid (PA) catabolic genes are up-regulated during growth in SCFM and are required for full virulence in a Caenorhabditis elegans host model. In this work, we investigated the role of phenylalanine, one of the aromatic amino acids present in SCFM, as an inducer of the PA catabolic pathway. Phenylalanine degradation intermediates were used as sole carbon sources for growth and gene reporter experiments. In addition to phenylalanine and PA, phenylethylamine, phenylpyruvate, and 2-phenylacetamide were usable as sole carbon sources by wild type B. cenocepacia K56-2, but not by a PA catabolism-defective mutant. EMSA analysis showed that the binding of PaaR, the negative regulator protein of B. cenocepacia PA catabolism, to PA regulatory DNA could only be relieved by phenylacetyl-Coenzyme A (PA-CoA), but not by any of the putative phenylalanine degradation intermediates. Taken together, our results show that in B. cenocepacia, phenylalanine is catabolized to PA and induces PA catabolism through PA activation to PA-CoA. Thus, PaaR shares the same inducer with PaaX, the regulator of PA catabolism in Escherichia coli, despite belonging to a different protein family.

  12. An Early Morning Sputum Sample Is Necessary for the Diagnosis of Pulmonary Tuberculosis, Even with More Sensitive Techniques: A Prospective Cohort Study among Adolescent TB-Suspects in Uganda

    PubMed Central

    Kateete, David P.; Wajja, Anne; Bugumirwa, Eric; Mboowa, Gerald; Namaganda, Carolyn; Nakayita, Germine; Nassolo, Maria; Mumbowa, Francis; Asiimwe, Benon B.; Waako, James; Verver, Suzanne; Musoke, Philippa; Mayanja-Kizza, Harriet; Joloba, Moses L.

    2012-01-01

    The World Health Organization (WHO) recommends collection of two sputum samples for tuberculosis (TB) diagnosis, with at least one being an early morning (EM) using smear microscopy. It remains unclear whether this is necessary even when sputum culture is employed. Here, we determined the diagnostic yield from spot and the incremental yield from the EM sputum sample cultures among TB-suspected adolescents from rural Uganda. Sputum samples (both spot and early-morning) from 1862 adolescents were cultured by the Lowenstein-Jensen (LJ) and Mycobacterium Growth Indicator Tube (MGIT) methods. For spot samples, the diagnostic yields for TB were 19.0% and 57.1% with LJ and MGIT, respectively, whereas the incremental yields (not totals) of the early-morning sample were 9.5% and 42.9% (P < 0.001) with LJ and MGIT, respectively. Among TB-suspected adolescents in rural Uganda, the EM sputum culture has a high incremental diagnostic yield. Therefore, EM sputum in addition to spot sample culture is necessary for improved TB case detection. PMID:23304491

  13. Two liquid medium systems, mycobacteria growth indicator tube and MB redox tube, for Mycobacterium tuberculosis isolation from sputum specimens.

    PubMed

    Heifets, L; Linder, T; Sanchez, T; Spencer, D; Brennan, J

    2000-03-01

    Two manual liquid medium systems, the Mycobacteria Growth Indicator Tube (MGIT) and MB Redox tube systems, were evaluated in comparison to the radiometric BACTEC-460 semiautomated system for recovery of Mycobacterium tuberculosis from sputum specimens. The highest level of recovery, from a total of 77 culture-positive specimens, occurred with the BACTEC-460 system (92.2%), followed by the MB Redox tube (80.5%) and the MGIT (63.6%) systems. The shortest time to detection was observed also among the cultures in BACTEC-460: a mean of 12 days to a growth index (GI) of 10 and 15 days to a GI of 500. The mean times for the other systems were 16 days for the MB Redox tube system and 17.4 days for the MGIT system. The proportion of cultures grown after more than 3 weeks of incubation was only 2.8 or 8.4% in BACTEC-460 (for a GI of 10 or 500) but 17.7% in MB Redox and 22.5% in MGIT. Despite these differences in comparison to the BACTEC-460 system and some differences between the MGIT and MB Redox tube systems, either of the two manual liquid medium systems presents a reasonable alternative to the BACTEC-460 system, especially for laboratories with a limited workload, and a valuable element in the laboratory protocol, in conjunction with solid media, for obtaining rapid detection of growth from about 80% of culture-positive specimens and for better overall recovery of M. tuberculosis. PMID:10699027

  14. [Pharmacological effects of N-acetyl-L-cysteine on the respiratory tract. (I). Quantitative and qualitative changes in respiratory tract fluid and sputum (author's transl)].

    PubMed

    Kogi, K; Saito, T; Kasé, Y; Hitoshi, T

    1981-06-01

    The following three experiments were performed to determine the effects of N-acetyl-L-cysteine (NAC) on the quantity and quality of respiratory tract fluid (RTF) and sputum. All drugs used were administered into the stomach through a gastric tube. 1) Indirect measurement of bronchial secretion in rats, which was expressed by the amounts of dye excreted into the respiratory tract, was carried out according the the Sakuno's method, with some modification. Some expectorants of the secretomotor type, such as bromhexine and pilocarpine, significantly increased the secretion, even at low doses. On the other hand, mucolytic agents such as NAC augmented the secretion only in doses of 500 to 1500 mg/kg. 2)As a direct method of measurements, Kasé's modification of Perry and Boyd's method was used to collect RTF, quantitatively, from rabbits. The RTF of healthy rabbits was colorless and watery. The administration of NAC in doses of 500 to 1500 mg/kg augmented the output volume and RTF became slightly turbid, probably due to an increase in the viscous mucus. 3) Rabbits with subacute bronchitis were prepared by long-term exposure to air contaminated with SO2 gas and sputa were collected before and after administration of NAC, respectively, according to the Kase's method. The sputa were opalescent and viscous gel included nodular masses. The administration of NAC, 1000 and 1500 mg/kg resulted in a dose dependent decrease in the relative viscosity. The percent-decreased in viscosity with NAC was statistically correlated with that in amounts of dry matter, those in protein and polysaccharide in the sputa. From the results described above, it was concluded that NAC given into the stomach can liquefy sputum by splitting mucoprotein disulphide linkages, that is, altering the rheological characteristics of sputum to facilitate expectoration. PMID:7286849

  15. [Pharmacological effects of N-acetyl-L-cysteine on the respiratory tract. (I). Quantitative and qualitative changes in respiratory tract fluid and sputum (author's transl)].

    PubMed

    Kogi, K; Saito, T; Kasé, Y; Hitoshi, T

    1981-06-01

    The following three experiments were performed to determine the effects of N-acetyl-L-cysteine (NAC) on the quantity and quality of respiratory tract fluid (RTF) and sputum. All drugs used were administered into the stomach through a gastric tube. 1) Indirect measurement of bronchial secretion in rats, which was expressed by the amounts of dye excreted into the respiratory tract, was carried out according the the Sakuno's method, with some modification. Some expectorants of the secretomotor type, such as bromhexine and pilocarpine, significantly increased the secretion, even at low doses. On the other hand, mucolytic agents such as NAC augmented the secretion only in doses of 500 to 1500 mg/kg. 2)As a direct method of measurements, Kasé's modification of Perry and Boyd's method was used to collect RTF, quantitatively, from rabbits. The RTF of healthy rabbits was colorless and watery. The administration of NAC in doses of 500 to 1500 mg/kg augmented the output volume and RTF became slightly turbid, probably due to an increase in the viscous mucus. 3) Rabbits with subacute bronchitis were prepared by long-term exposure to air contaminated with SO2 gas and sputa were collected before and after administration of NAC, respectively, according to the Kase's method. The sputa were opalescent and viscous gel included nodular masses. The administration of NAC, 1000 and 1500 mg/kg resulted in a dose dependent decrease in the relative viscosity. The percent-decreased in viscosity with NAC was statistically correlated with that in amounts of dry matter, those in protein and polysaccharide in the sputa. From the results described above, it was concluded that NAC given into the stomach can liquefy sputum by splitting mucoprotein disulphide linkages, that is, altering the rheological characteristics of sputum to facilitate expectoration.

  16. Direct Detection of Rifampin- and Isoniazid-Resistant Mycobacterium tuberculosis in Auramine-Rhodamine-Positive Sputum Specimens by Real-Time PCR

    PubMed Central

    Ruiz, Maite; Torres, Maria J.; Llanos, Ana C.; Arroyo, Aurelio; Palomares, Jose C.; Aznar, Javier

    2004-01-01

    Our objective was to evaluate the feasibility of a molecular assay based on a real-time PCR technique, carried out with a LightCycler instrument (Roche Biochemicals), to identify Mycobacterium tuberculosis bacilli and to detect rifampin and isoniazid resistance in DNA extracts from sputum samples. We studied three genes: rpoB, which is associated with rifampin resistance, and katG and inhA, which are associated with isoniazid resistance. A total of 205 sputum samples collected from 108 patients diagnosed with pulmonary tuberculosis with positive auramine-rhodamine-staining (AR) sputum samples, were tested. The sensitivities of the LightCycler PCR assay for the positive AR specimens was 97.5% (200 of 205) for rpoB and inhA genes and 96.5% (198 of 205) for the katG gene. For the total number of patients tested, the sensitivity was 100% (108 of 108 patients) for rifampin, whereas the sensitivity was 98.1% (106 of 108 patients) for isoniazid. Full agreement was found with the Bactec MGIT 960 method and the genotype inferred from the LightCycler data for rifampin. The phenotypic method for isoniazid reported 13 resistant strains (≥0.1 μg/ml). In seven (53.8%) strains there was a concordance between both methods, but we found that six (46.2%) strains reported as resistant by the phenotypic method were determined to be susceptible by real-time PCR. For the 75 strains reported as susceptible by the phenotypic method, the concordance with the LightCycler data was 100%. Our results demonstrate that rifampin-resistant M. tuberculosis could be detected in DNA extracted from auramine-rhodamine-positive sputum samples in a single-tube assay that took less than 3 h to perform for a collection of auramine-rhodamine-positive specimens obtained from patients with culture-documented pulmonary tuberculosis. Similarly, this occurs in half of the isoniazid-resistant M. tuberculosis DNA extracted from auramine-rhodamine-positive specimens. PMID:15071008

  17. Determination of ciprofloxacin and levofloxacin in human sputum collected from cystic fibrosis patients using microextraction by packed sorbent-high performance liquid chromatography photodiode array detector.

    PubMed

    Locatelli, Marcello; Ciavarella, Maria Teresa; Paolino, Donatella; Celia, Christian; Fiscarelli, Ersilia; Ricciotti, Gabriella; Pompilio, Arianna; Di Bonaventura, Giovanni; Grande, Rossella; Zengin, Gokhan; Di Marzio, Luisa

    2015-11-01

    This paper reports a new, easy, cheap, and fast MEPS-HPLC-PDA method for the simultaneous analysis of ciprofloxacin and levofloxacin, two fluoroquinolones (FLQs) commonly used for the treatment of pulmonary infections in cystic fibrosis (CF) patients. The FLQs were resolved on a Discovery C8 column (250mm×4.6mm; 5μm particle size) using an isocratic elution with a run time of 15min, without further purification. The method was validated over concentrations ranging from 0.05 to 2μg/mL for both analytes in human sputum, and enrofloxacin was used as internal standard. This method was successfully tested to detect FLQs in sputum collected from CF patients. The MEPS-HPLC-PDA method was validated using biological samples collected from CF patients orally or intravenously injected with FLQs. The resultant data showed that the method is selective, sensitive and robust over range of concentrations for both FLQs. The limit of quantification of the method was 0.05μg/mL for both analytes (comparable to more complex and expensive instrument configurations), weighted-matrix-matched standard curves showed a good linearity up to 2μg/mL, and parallelism tests were also successfully assessed. The intra- and inter-day precision (RSD%) values were ≤10.4% and ≤11.1%, respectively, for all range of analysis. The intra- and inter-day trueness (Bias%) values are ranged from -11.8% to 7.25% for both antibiotic drugs. At the best of our knowledge, this is the first MEPS-HPLC-PDA based method that uses MEPS procedure for simultaneous determination of ciprofloxacin and levofloxacin in human sputum. The method was tested successfully on real sputum samples by following a conventional drug administration. Furthermore, the MEPS-HPLC-PDA based method provides more advantages to detect and analyze quickly the antibiotic drugs in biological matrices than other analytical procedures reported in literature.

  18. The Conservation and Application of Three Hypothetical Protein Coding Gene for Direct Detection of Mycobacterium tuberculosis in Sputum Specimens

    PubMed Central

    Qin, Lianhua; Gao, Shihui; Wang, Jie; Zheng, Ruijuan; Lu, Junmei; Hu, Zhongyi

    2013-01-01

    Background Accurate and early diagnosis of tuberculosis (TB) is of major importance in the control of TB. One of the most important technical advances in diagnosis of tuberculosis is the development of nucleic acid amplification (NAA) tests. However, the choice of the target sequence remains controversial in NAA tests. Recently, interesting alternatives have been found in hypothetical protein coding sequences from mycobacterial genome. Methodology/Principal Findings To obtain rational biomarker for TB diagnosis, the conservation of three hypothetical genes was firstly evaluated in 714 mycobacterial strains. The results showed that SCAR1 (Sequenced Characterized Amplified Region) based on Rv0264c coding gene showed the highest conservation (99.8%) and SCAR2 based on Rv1508c gene showed the secondary high conservation (99.7%) in M. tuberculosis (MTB) strains. SCAR3 based on Rv2135c gene (3.2%) and IS6110 (8%) showed relatively high deletion rate in MTB strains. Secondly, three SCAR markers were evaluated in 307 clinical sputum from patients in whom TB was suspected or patients with diseases other than TB. The amplification of IS6110 and 16SrRNA sequences together with both clinical and bacteriological identification was as a protocol to evaluate the efficacy of SCAR markers. The sensitivities and specificities, positive predictive value (PPV) and negative predictive value (NPV) of all NAA tests were higher than those of bacteriological detection. In four NAA tests, IS6110 and SCAR3 showed the highest PPV (100%) and low NPV (70% and 68.8%, respectively), and SCAR1 and SCAR2 showed the relatively high PPV and NPV (97% and 82.6%, 95.6% and 88.8%, respectively). Conclusions/Significance Our result indicated that SCAR1 and SCAR2 with a high degree of sequence conservation represent efficient and promising alternatives as NAA test targets in identification of MTB. Moreover, the targets developed from this study may provide more alternative targets for the development of a

  19. Sputum stain for mycobacteria

    MedlinePlus

    Murray PR. The clinician and the microbiology laboratory. In: Bennett JE, Dolin R, Blaser MJ, eds. Mandell, Douglass, and Bennett's Principles and Practice of Infectious Diseases . 6th ed. Philadelphia, ...

  20. Sputum Gram stain

    MedlinePlus

    ... under a microscope, checking for bacteria and white blood cells. The color, size, and shape of the cells help identify ... material that has a foul odor or unusual color. The test may also ... other signs and symptoms of respiratory disease or infection.

  1. Microbial effects on sorption and transport of actinides in tuff samples from the Nevada Test Site and soils from McGuire AFB, NJ

    NASA Astrophysics Data System (ADS)

    Fisher, J. C.; Gostic, R.; Gostic, J.; Czerwinski, K.; Moser, D. P.

    2009-12-01

    The sorption and behavior of various actinides were examined for two sets of environmental samples. The Nevada Test Site (NTS) harbors a variety of radionuclides resulting from atomic weapons testing from the 1950s-1990s. Modeling the transport of radionuclides at the NTS is difficult because each detonation cavity is a unique environment with distinct hydrologic characteristics, chemical composition, and microbial community structure. McGuire AFB was the site of an explosion that resulted in the burning of a BOMARC nuclear missile and deposition of particles containing high-fired oxides of Am, Pu, and U in soils on the base. Analysis of the NTS samples focused on sorption/desorption of 233-U and 241-Am in the presence/absence of bacteria, and work on the BOMARC cores addressed the potential role of microorganisms in mediating particle degradation and movement. Batch experiments with various NTS tuff samples and strains of bacteria showed that sorption of actinides may be enhanced by >25% under certain conditions by bacteria. Sorption of 233-U was highly dependent on carbonate concentrations in the liquid matrix, while 241-Am was unaffected. Different bacterial species also affected sorption differently. Sorption kinetics for both actinides were rapid, with maximum sorption usually occurring within 4 hours. Actinides bound tightly to tuff and little desorption occurred in carbonate-free batch experiments. Column experiments showed that bacterial cultures in minimal salts buffer desorbed significantly more 233-U from tuff than low carbonate NTS water, but less than 30 mM bicarbonate buffer. Hot particles in the BOMARC cores were located using CT mapping and were extracted from the soil prior to analysis of core sections by gamma spectroscopy. Subcores for DNA extraction and culturing were collected from soil in direct contact with hot particles. The extracted particles consisted of a mixture of weapons-grade Pu, 241-Am and 235-U and ranged in activity from 5-66 k

  2. Assessment of the quantity of microorganisms associated with bronchiectasis in saliva, sputum and nasal lavage after periodontal treatment: a study protocol of a randomised controlled trial

    PubMed Central

    Pinto, Erika Horácio; Longo, Priscila Larcher; de Camargo, Caroline Cristina Batista; Dal Corso, Simone; Lanza, Fernanda De Cordoba; Stelmach, Rafael; Athanazio, Rodrigo; Fernandes, Kristianne Porta Santos; Mayer, Marcia Pinto Alves; Bussadori, Sandra Kalil; Mesquita Ferrari, Raquel Agnelli; Horliana, Anna Carolina Ratto Tempestini

    2016-01-01

    Introduction The association between periodontal disease (PD) and chronic obstructive pulmonary disease (COPD) has been widely studied, with aspiration of periodontal pathogens being one of the most accepted causal mechanisms for pulmonary exacerbation. Periodontal treatment (PT) was associated with a decrease in these exacerbations. Bronchiectasis is a pulmonary disease that has many similarities to COPD; however, there are no studies correlating this condition to PD thus far. This study will evaluate if PT reduces proinflammatory cytokines in serum and saliva, as well as halitosis and the amount of microorganisms associated with exacerbation of bronchiectasis in saliva, sputum and nasal lavage 3 months after PT. Methods and analysis A total of 182 patients with PD and bronchiectasis will be randomly allocated to group 1 (positive control; scaling and root planing (SRP)+oral hygiene (OH)) or group 2 (experimental; SRP+photodynamic therapy+OH). After 3 months, samples of saliva, nasal lavage and sputum will be collected to determine the level of Pseudomonas aeruginosa, Staphylococcus aureus and Porphyromonas gingivalis by quantitative PCR. This protocol will determine the efficacy of PT in reducing the most likely niches of bronchiectasis exacerbation by comparing pre- and post-treatment microbiology samples. Furthermore, there will be assessment of oral halitosis and verification of inflammatory cytokines in serum and saliva. Ethics and dissemination This protocol has been approved by the Research Ethics Committee of Universidade Nove de Julho. Data will be published in a peer-reviewed journal. Trial registration number NCT02514226. PMID:27084279

  3. Active Case Finding of Pulmonary Tuberculosis through Screening of Respiratory Symptomatics Using Sputum Microscopy: Is It Time to Change the Paradigm?

    PubMed Central

    del Portillo-Mustieles, Eva Carolina; Laniado-Laborín, Rafael

    2013-01-01

    Background. One of the main strategies for the early detection of pulmonary tuberculosis (PTB) is through the screening of individuals with symptoms compatible with PTB. Although this is programmatic strategy for active case finding, its yield is not well known. Objective. To determine the yield of pulmonary tuberculosis active case finding through the screening of respiratory symptomatic (RS) patients at a general hospital. Methods. RS patients were defined as subjects complaining of cough and/or sputum for a period of 2 or more weeks. Outpatients and their companions were approached while they waited in the outpatient care areas of the hospital to detect RS. Two samples from different days or 2 samples taken 2 hours apart on the same day were collected. Results. 122 RS patients were identified. Fifty-seven patients (46.7%) had at least one sputum sample analyzed. Three patients presented a positive smear and 2 were culture positive; neither had upper airway symptoms. None of the patients with productive cough and upper airway symptoms had a positive smear (P = 0.07). Only 19 (33.3%) returned to the laboratory to retrieve their results. Conclusion. Current strategy to screen RS patients based only on clinical data has a low compliance. Specific strategies to increase compliance (removal of barriers, incentives, etc.) should be implemented. PMID:23533747

  4. Rapid detection of Mycobacterium tuberculosis complex by real-time PCR in sputum samples and its use in the routine diagnosis in a reference laboratory.

    PubMed

    Watanabe Pinhata, Juliana Maira; Cergole-Novella, Maria Cecilia; Moreira dos Santos Carmo, Andreia; Ruivo Ferro e Silva, Regina; Ferrazoli, Lucilaine; Tavares Sacchi, Claudio; Siqueira de Oliveira, Rosangela

    2015-09-01

    Tuberculosis (TB) is an infectious disease of global distribution, constituting a serious public health problem in Brazil. São Paulo State, located in the south-east of Brazil, notified 16,580 new TB cases in 2013. The Instituto Adolfo Lutz is a public health reference laboratory for TB diagnosis for all the State. Considering that rapid and accurate diagnosis is essential for TB control, the aim of this study was to evaluate the use of an in-house real-time (RT)-PCR assay targeting the mpt64 gene in the routine diagnosis of TB, and to compare this technique with smear microscopy and culture. From August 2012 to October 2013, 715 sputum samples from 657 patients were included in the study. Smear microscopy, culture, phenotypic and PRA-hsp65 identification of mycobacteria, and mpt64 RT-PCR were performed. With respect to confirmed TB cases (n = 62/657; 9.4%), smear microscopy had a sensitivity of 82.3%. Culture and RT-PCR showed the same sensitivity, i.e. 90.3%. Specificity was 99.7, 99.4 and 98.6% for smear microscopy, culture and RT-PCR, respectively. mpt64 RT-PCR showed high sensitivity and specificity for the detection of Mycobacterium tuberculosis complex in sputum samples. This technique can be deployed in laboratories that do not have a rapid test for TB available, enabling the performance of TB diagnosis in up to 5 h.

  5. Physiological levels of nitrate support anoxic growth by denitrification of Pseudomonas aeruginosa at growth rates reported in cystic fibrosis lungs and sputum.

    PubMed

    Line, Laura; Alhede, Morten; Kolpen, Mette; Kühl, Michael; Ciofu, Oana; Bjarnsholt, Thomas; Moser, Claus; Toyofuku, Masanori; Nomura, Nobuhiko; Høiby, Niels; Jensen, Peter Ø

    2014-01-01

    Chronic Pseudomonas aeruginosa lung infection is the most severe complication in patients with cystic fibrosis (CF). The infection is characterized by the formation of biofilm surrounded by numerous polymorphonuclear leukocytes (PMNs) and strong O2 depletion in the endobronchial mucus. We have reported that O2 is mainly consumed by the activated PMNs, while O2 consumption by aerobic respiration is diminutive and nitrous oxide (N2O) is produced in infected CF sputum. This suggests that the reported growth rates of P. aeruginosa in lungs and sputum may result from anaerobic respiration using denitrification. The growth rate of P. aeruginosa achieved by denitrification at physiological levels (~400 μM) of nitrate (NO(-) 3) is however, not known. Therefore, we have measured growth rates of anoxic cultures of PAO1 and clinical isolates (n = 12) in LB media supplemented with NO(-) 3 and found a significant increase of growth when supplementing PAO1 and clinical isolates with ≥150 μM NO(-) 3 and 100 μM NO(-) 3, respectively. An essential contribution to growth by denitrification was demonstrated by the inability to establish a significantly increased growth rate by a denitrification deficient ΔnirS-N mutant at <1 mM of NO(-) 3. Activation of denitrification could be achieved by supplementation with as little as 62.5 μM of NO(-) 3 according to the significant production of N2O by the nitrous oxide reductase deficient ΔnosZ mutant. Studies of the promoter activity, gene transcripts, and enzyme activity of the four N-oxide reductases in PAO1 (Nar, Nir, Nor, Nos) further verified the engagement of denitrification, showing a transient increase in activation and expression and rapid consumption of NO(-) 3 followed by a transient increase of NO(-) 2. Growth rates obtained by denitrification in this study were comparable to our reported growth rates in the majority of P. aeruginosa cells in CF lungs and sputum. Thus, we have demonstrated that denitrification is required

  6. Effects of Low-Dose and Long-Term Treatment with Erythromycin on Interleukin-17 and Interleukin-23 in Peripheral Blood and Induced Sputum in Patients with Stable Chronic Obstructive Pulmonary Disease

    PubMed Central

    Tan, Caimei; Huang, Huijuan; Zhang, Jianquan; He, Zhiyi; Zhong, Xiaoning; Bai, Jing

    2016-01-01

    Objective. To study the effects of low-dose and long-term treatment with erythromycin on IL-17 and IL-23, in peripheral blood and induced sputum, in patients with stable chronic obstructive pulmonary disease (COPD). Methods. Patients were randomly divided into placebo-treated group, group A (12 months of additive treatment with erythromycin, N = 18), and group B (6 months of additive treatment with erythromycin followed by 6 months of follow-up, N = 18). Inflammatory cells in induced sputum, pulmonary function, and the 6-minute walk distance (6MWD) were analyzed. Concentrations of IL-17 and IL-23 in peripheral blood and sputum were measured using enzyme-linked immunosorbent assays. Results. After treatment, sputum and peripheral blood concentrations of IL-17 and IL-23 significantly decreased in groups A and B compared with placebo-treated group. There were no significant differences after erythromycin withdrawal at months 9 and 12 in group B compared with placebo-treated group. An increase in 6MWD was observed after treatment. Conclusions. Erythromycin was beneficial and reduced airway inflammation in COPD patients. Underlying mechanisms may involve inhibition of IL-17 and IL-23 mediated airway inflammation. COPD patients treated with erythromycin for 6 months experienced improved exercise capacity. Finally, treatment for 12 months may be more effective than treatment for 6 months. PMID:27127346

  7. Impact of the Mycobaterium africanum West Africa 2 Lineage on TB Diagnostics in West Africa: Decreased Sensitivity of Rapid Identification Tests in The Gambia

    PubMed Central

    Ofori-Anyinam, Boatema; Kanuteh, Fatoumatta; Okoi, Catherine; Dolganov, Gregory; Schoolnik, Gary; Secka, Ousman; Antonio, Martin; de Jong, Bouke C.; Gehre, Florian

    2016-01-01

    Background MPT64 rapid speciation tests are increasingly being used in diagnosis of tuberculosis (TB). Mycobacterium africanum West Africa 2 (Maf 2) remains an important cause of TB in West Africa and causes one third of disease in The Gambia. Since the introduction of MPT64 antigen tests, a higher than expected rate of suspected non-tuberculous mycobacteria (NTM) was seen among AFB smear positive TB suspects, which led us to prospectively assess sensitivity of the MPT64 antigen test in our setting. Methodology/Principal Findings We compared the abundance of mRNA encoded by the mpt64 gene in sputa of patients with untreated pulmonary TB caused by Maf 2 and Mycobacterium tuberculosis (Mtb). Subsequently, prospectively collected sputum samples from presumptive TB patients were inoculated in the BACTEC MGIT 960 System. One hundred and seventy-three acid fast bacilli (AFB)-positive and blood agar negative MGIT cultures were included in the study. Cultures were tested on the day of MGIT positivity with the BD MGIT TBc Identification Test. A random set of positives and all negatives were additionally tested with the SD Bioline Ag MPT64 Rapid. MPT64 negative cultures were further incubated at 37°C and retested until positive. Bacteria were spoligotyped and assigned to different lineages. Maf 2 isolates were 2.52-fold less likely to produce a positive test result and sensitivity ranged from 78.4% to 84.3% at the beginning and end of the recommended 10 day testing window, respectively. There was no significant difference between the tests. We further showed that the decreased rapid test sensitivity was attributable to variations in mycobacterial growth behavior and the smear grades of the patient. Conclusions/Significance In areas where Maf 2 is endemic MPT64 tests should be cautiously used and MPT64 negative results confirmed by a second technique, such as nucleic acid amplification tests, to avoid their misclassification as NTMs. PMID:27387550

  8. Analysis of Lung Microbiota in Bronchoalveolar Lavage, Protected Brush and Sputum Samples from Subjects with Mild-To-Moderate Cystic Fibrosis Lung Disease.

    PubMed

    Hogan, Deborah A; Willger, Sven D; Dolben, Emily L; Hampton, Thomas H; Stanton, Bruce A; Morrison, Hilary G; Sogin, Mitchell L; Czum, Julianna; Ashare, Alix

    2016-01-01

    Individuals with cystic fibrosis (CF) often acquire chronic lung infections that lead to irreversible damage. We sought to examine regional variation in the microbial communities in the lungs of individuals with mild-to-moderate CF lung disease, to examine the relationship between the local microbiota and local damage, and to determine the relationships between microbiota in samples taken directly from the lung and the microbiota in spontaneously expectorated sputum. In this initial study, nine stable, adult CF patients with an FEV1>50% underwent regional sampling of different lobes of the right lung by bronchoalveolar lavage (BAL) and protected brush (PB) sampling of mucus plugs. Sputum samples were obtained from six of the nine subjects immediately prior to the procedure. Microbial community analysis was performed on DNA extracted from these samples and the extent of damage in each lobe was quantified from a recent CT scan. The extent of damage observed in regions of the right lung did not correlate with specific microbial genera, levels of community diversity or composition, or bacterial genome copies per ml of BAL fluid. In all subjects, BAL fluid from different regions of the lung contained similar microbial communities. In eight out of nine subjects, PB samples from different regions of the lung were also similar in microbial community composition, and were similar to microbial communities in BAL fluid from the same lobe. Microbial communities in PB samples were more diverse than those in BAL samples, suggesting enrichment of some taxa in mucus plugs. To our knowledge, this study is the first to examine the microbiota in different regions of the CF lung in clinically stable individuals with mild-to-moderate CF-related lung disease. PMID:26943329

  9. Exposing the Three-Dimensional Biogeography and Metabolic States of Pathogens in Cystic Fibrosis Sputum via Hydrogel Embedding, Clearing, and rRNA Labeling

    PubMed Central

    DePas, William H.; Starwalt-Lee, Ruth; Van Sambeek, Lindsey; Ravindra Kumar, Sripriya

    2016-01-01

    ABSTRACT Physiological resistance to antibiotics confounds the treatment of many chronic bacterial infections, motivating researchers to identify novel therapeutic approaches. To do this effectively, an understanding of how microbes survive in vivo is needed. Though much can be inferred from bulk approaches to characterizing complex environments, essential information can be lost if spatial organization is not preserved. Here, we introduce a tissue-clearing technique, termed MiPACT, designed to retain and visualize bacteria with associated proteins and nucleic acids in situ on various spatial scales. By coupling MiPACT with hybridization chain reaction (HCR) to detect rRNA in sputum samples from cystic fibrosis (CF) patients, we demonstrate its ability to survey thousands of bacteria (or bacterial aggregates) over millimeter scales and quantify aggregation of individual species in polymicrobial communities. By analyzing aggregation patterns of four prominent CF pathogens, Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus sp., and Achromobacter xylosoxidans, we demonstrate a spectrum of aggregation states: from mostly single cells (A. xylosoxidans), to medium-sized clusters (S. aureus), to a mixture of single cells and large aggregates (P. aeruginosa and Streptococcus sp.). Furthermore, MiPACT-HCR revealed an intimate interaction between Streptococcus sp. and specific host cells. Lastly, by comparing standard rRNA fluorescence in situ hybridization signals to those from HCR, we found that different populations of S. aureus and A. xylosoxidans grow slowly overall yet exhibit growth rate heterogeneity over hundreds of microns. These results demonstrate the utility of MiPACT-HCR to directly capture the spatial organization and metabolic activity of bacteria in complex systems, such as human sputum. PMID:27677788

  10. Evaluation of real-time loop-mediated isothermal amplification (RealAmp) for rapid detection of Mycobacterium tuberculosis from sputum samples.

    PubMed

    Li, Yiming; Shi, Lei; Pan, Anqi; Cao, Weiwei; Chen, Xun; Meng, Hecheng; Yan, He; Miyoshi, Shin-ichi; Ye, Lei

    2014-09-01

    Tuberculosis (TB) caused by Mycobacterium tuberculosis (MTB) leads to serious health problems as a chronic respiratory infectious disease. Here we established a real-time fluorescence loop-mediated isothermal amplification assay (RealAmp) using a portable ESE Quant tube scanner as a convenient rapid detection method for MTB. The method efficacy from sputum samples was further investigated, and the reaction time was only 20min with the detection limit low to 10(2)CFU/ml concentration of MTB. We assessed a total of 1067 samples by the RealAmp assay, comparing the results with smear microscopy and conventional culture methods. To examine whether the failure to detect TB by culturing is due to low sensitivity or true absence, we examined the culture negative samples by commercial real time PCR MTB detection kit, and the results were compared with RealAmp. The data showed that RealAmp assay had a higher positive rate than that of sputum smear and culture methods. RealAmp had a sensitivity of 96.70% and a specificity of 91.55% when compared with culture. In addition, its sensitivity and specificity were 95.29% and 86.88% respectively compared with examination of smear samples using light microscopy. The sensitivity of RealAmp in comparison to real time PCR was 98.25% and specificity was 99.11% in validation of culture negative samples. The present study revealed the newly established RealAmp assay as a convenient, efficient, sensitive and specific method that could be an alternative for rapid detection of MTB and a tool to validate culture and smear negative samples. Furthermore, the portability of the ESE Quant tube scanner also contributed to the promising application for grassroots and field detection of MTB.

  11. Analysis of Lung Microbiota in Bronchoalveolar Lavage, Protected Brush and Sputum Samples from Subjects with Mild-To-Moderate Cystic Fibrosis Lung Disease

    PubMed Central

    Hogan, Deborah A.; Willger, Sven D.; Dolben, Emily L.; Hampton, Thomas H.; Stanton, Bruce A.; Morrison, Hilary G.; Sogin, Mitchell L.; Czum, Julianna; Ashare, Alix

    2016-01-01

    Individuals with cystic fibrosis (CF) often acquire chronic lung infections that lead to irreversible damage. We sought to examine regional variation in the microbial communities in the lungs of individuals with mild-to-moderate CF lung disease, to examine the relationship between the local microbiota and local damage, and to determine the relationships between microbiota in samples taken directly from the lung and the microbiota in spontaneously expectorated sputum. In this initial study, nine stable, adult CF patients with an FEV1>50% underwent regional sampling of different lobes of the right lung by bronchoalveolar lavage (BAL) and protected brush (PB) sampling of mucus plugs. Sputum samples were obtained from six of the nine subjects immediately prior to the procedure. Microbial community analysis was performed on DNA extracted from these samples and the extent of damage in each lobe was quantified from a recent CT scan. The extent of damage observed in regions of the right lung did not correlate with specific microbial genera, levels of community diversity or composition, or bacterial genome copies per ml of BAL fluid. In all subjects, BAL fluid from different regions of the lung contained similar microbial communities. In eight out of nine subjects, PB samples from different regions of the lung were also similar in microbial community composition, and were similar to microbial communities in BAL fluid from the same lobe. Microbial communities in PB samples were more diverse than those in BAL samples, suggesting enrichment of some taxa in mucus plugs. To our knowledge, this study is the first to examine the microbiota in different regions of the CF lung in clinically stable individuals with mild-to-moderate CF-related lung disease. PMID:26943329

  12. The effect on sound generation of varying both gas flow rate and the viscosity of sputum-like gel in a simple tubular model.

    PubMed

    Jones, A Y; Jones, R D; Kwong, K; Burns, Y

    2000-01-01

    Gas flows of 2, 3, and 4 L/min were directed through a sputum-like gel with viscosities of 100, 150, and 200 P and placed in a tube similar in diameter to a human segmental bronchus (4 mm), which was immersed in a bath of water. The sound produced by gas flow through the gel was recorded with a hydrophone. Sound data were subjected to time-expanded waveforms and fast Fourier transform (FFT) analysis. This study demonstrated that the number of crackles generated was directly related to the flow rate and inversely related to gel viscosity. The initial deflection width (IDW), two-cycle duration (2 CD), and peak-to-peak amplitude of crackles were significantly affected by the gas flow rate but not the viscosity of the gel. A lower gas flow rate generated crackles with longer IDW and 2 CD, but higher gas flow rates generated crackles with higher amplitude. Peak sound intensity measured from FFT increased as flow rate increased but decreased as the viscosity of the gel increased. At low gas flows, no gel-induced crackle sound was generated within the data capture window when the most viscous gel was examined. A digital video image of gas flow through the gel was captured, and this confirmed the absence of bubbles or slug formation at low flows through 200 P gel during the 3 seconds of data acquisition. This study describes some characteristics of crackles generated from different combinations of gas flow and gel viscosity and suggests that "coarse crackles" results from the explosion of gas bubbles in pulmonary secretions. Health care practitioners should consider the combined effect of rate of inspiratory gas flow and sputum viscosity during auscultation of patients' lungs.

  13. Can Follow-Up Examination of Tuberculosis Patients Be Simplified? A Study in Chhattisgarh, India

    PubMed Central

    Kundu, Debashish; M. V. Kumar, Ajay; Satyanarayana, Srinath; Dewan, Puneet Kumar; Achuthan Nair, Sreenivas; Khaparde, Kshitij; Nayak, Priyakanta; Van den Bergh, Rafael; Manzi, Marcel; Enarson, Donald A.; Deshpande, Madhav Rao; Chandraker, Sachin

    2012-01-01

    Background Each follow-up during the course of tuberculosis treatment currently requires two sputum examinations. However, the incremental yield of the second sputum sample during follow-up of different types of tuberculosis patients has never been determined precisely. Objectives To assess the incremental yield of the second sputum sample in the follow-up of tuberculosis patients under the Revised National Tuberculosis Control Programme (RNTCP) in Chhattisgarh, India. Methodology A record review of tuberculosis (TB) patients registered in 2009 using a structured proforma from two sources, Tuberculosis and Laboratory Register, was undertaken in the six districts of Chhattisgarh, India. Results In smear positive cases, of 10,048 follow-up examinations, 45 (0.5%) were found to be smear positive only on the second sputum when the result of the first sample was negative. In smear negative pulmonary and extra pulmonary TB patients, of 6,206 follow-up smear examinations, 11(0.2%) were found to be smear positive. Conclusions The incremental yield of a second smear examination was very low, indicating that examination of one sputum sample is enough during follow-up among TB patients. There is insufficient yield to support sputum smear microscopy for monitoring smear negative pulmonary TB and extra pulmonary TB patients. These results indicate that the follow-up smear microscopy can be substantially simplified with favourable resource implications. PMID:23227230

  14. Impact of nonlinear interactions of pharmacokinetics and MICs on sputum bacillary kill rates as a marker of sterilizing effect in tuberculosis.

    PubMed

    Chigutsa, Emmanuel; Pasipanodya, Jotam G; Visser, Marianne E; van Helden, Paul D; Smith, Peter J; Sirgel, Frederick A; Gumbo, Tawanda; McIlleron, Helen

    2015-01-01

    The relationships between antituberculosis drug exposure and treatment effects on humans receiving multidrug therapy are complex and nonlinear. In patients on treatment, an analysis of the rate of decline in the sputum bacillary burden reveals two slopes. The first is the α-slope, which is thought to reflect bactericidal effect, followed by a β-slope, which is thought to reflect sterilizing activity. We sought to characterize the effects of standard first-line treatment on sterilizing activity. Fifty-four patients receiving combination therapy for pulmonary tuberculosis in a clinical trial had drug concentrations measured and Mycobacterium tuberculosis isolates available for MIC identification. Sputum sample cultures were performed at baseline and weekly for 8 weeks. A time-to-event model based on the days to positivity in the liquid cultures was used to estimate the β-slope. The pharmacokinetic parameters of rifampin, isoniazid, ethambutol, and pyrazinamide were determined for each patient. Multivariate adaptive regression splines analyses, which simultaneously perform linear and nonlinear analyses, were used to identify the relationships between the predictors and the β-slope. The potential predictors examined included HIV status, lung cavitation, 24-h area under the concentration-time curve (AUC), peak drug concentration (Cmax), AUC/MIC ratio, Cmax/MIC ratio, and the time that that concentration persisted above MIC. A rifampin Cmax of >8.2 mg/liter and a pyrazinamide AUC/MIC of >11.3 were key predictors of the β-slope and interacted positively to increase the β-slope. In patients with a rifampin AUC of <35.4 mg · h/liter, an increase in the pyrazinamide AUC/MIC and/or ethambutol Cmax/MIC increased the β-slope, while increasing isoniazid Cmax decreased it, suggesting isoniazid antagonism. Antibiotic concentrations and MICs interact in a nonlinear fashion as the main drivers of a sterilizing effect. The results suggest that faster speeds of sterilizing

  15. Geographic Differences in Time to Culture Conversion in Liquid Media: Tuberculosis Trials Consortium Study 28. Culture Conversion Is Delayed in Africa

    PubMed Central

    Mac Kenzie, William R.; Heilig, Charles M.; Bozeman, Lorna; Johnson, John L.; Muzanye, Grace; Dunbar, Denise; Jost, Kenneth C.; Diem, Lois; Metchock, Beverly; Eisenach, Kathleen; Dorman, Susan; Goldberg, Stefan

    2011-01-01

    Background Tuberculosis Trials Consortium Study 28, was a double blind, randomized, placebo-controlled, phase 2 clinical trial examining smear positive pulmonary Mycobacterium tuberculosis. Over the course of intensive phase therapy, patients from African sites had substantially delayed and lower rates of culture conversion to negative in liquid media compared to non-African patients. We explored potential explanations of this finding. Methods In TBTC Study 28, protocol-correct patients (n = 328) provided spot sputum specimens for M. tuberculosis culture in liquid media, at baseline and weeks 2, 4, 6 and 8 of study therapy. We compared sputum culture conversion for African and non-African patients stratified by four baseline measures of disease severity: AFB smear quantification, extent of disease on chest radiograph, cavity size and the number of days to detection of M. tuberculosis in liquid media using the Kaplan-Meier product-limit method. We evaluated specimen processing and culture procedures used at 29 study laboratories serving 27 sites. Results African TB patients had more extensive disease at enrollment than non-African patients. However, African patients with the least disease by the 4 measures of disease severity had conversion rates on liquid media that were substantially lower than conversion rates in non-African patients with the greatest extent of disease. HIV infection, smoking and diabetes did not explain delayed conversion in Africa. Some inter-site variation in laboratory processing and culture procedures within accepted practice for clinical diagnostic laboratories was found. Conclusions Compared with patients from non-African sites, African patients being treated for TB had delayed sputum culture conversion and lower sputum conversion rates in liquid media that were not explained by baseline severity of disease, HIV status, age, smoking, diabetes or race. Further investigation is warranted into whether modest variation in laboratory

  16. Diagnosing Lung Cancers through Examination of Micro-RNA Biomarkers in Blood, Plasma, Serum and Sputum: A Review and Summary of Current Literature

    PubMed Central

    Gyoba, Jennifer; Shan, Shubham; Roa, Wilson; Bédard, Eric L. R.

    2016-01-01

    Lung cancer is the leading cause of cancer related morbidity and mortality worldwide. Currently, the vast majority of lung cancers are diagnosed at a late stage, when patients become symptomatic leading to dismal, less than 15% five-year survival rates. Evidence has demonstrated that screening computed tomography scans can be used to detect lung cancer, but these scans have high false positive rates. Therefore, there is a continued need for the development of minimally-invasive methods to screen the high risk population and diagnose lung cancer at an earlier, curable stage. One such promising area is the use micro-RNAs. These are short, non-coding RNA molecules that have been shown in previous research to be dysregulated in cancers. This review will focus on the potential use of miRNA levels in various biological fluids (whole blood, plasma, serum, and sputum) and demonstrate their potential utility as screening and diagnostic biomarkers for lung cancer. Current research will be analyzed and compared, and future directions in establishing the use of miRNAs for detecting lung cancer will be discussed. PMID:27043555

  17. The late Greco-Roman and Byzantine contribution towards the evolution of laboratory examinations of bodily excrement. Part 2: sputum, vomit, blood, sweat, autopsies.

    PubMed

    Diamandopoulos, Athanasios A; Goudas, Pavlos C

    2005-01-01

    Although the establishment of medical laboratory institutions was a continuous process that matured only after the 16th century, several attempts had already been made to attain a diagnosis by investigating bodily excrement. In the first part of our work, published in a previous issue of this journal, we presented data on urine, sperm, menses and stools. In this paper we present data on sputum, vomit, blood, sweat, and autopsies, thus completing the list of human materials used for laboratory examinations. All the data used are extracted from codices of Late Antiquity and Byzantium and translated by us. We did not study medical texts from the other great ancestors of Western medicine, namely Arabic and Jewish writings. From the texts cited, it is apparent that the lack of technological means was no obstacle for the doctor to create an "examinational" mind, i.e., to try to correlate the macroscopic findings in the excrement with the pathophysiological mechanism that induced them, solely with the use of the senses. This not only applies to the examination of urine, as is commonly assumed, but also to many other excrements of the upper and lower orifices of the body, as well as the human body as a whole. PMID:15653449

  18. Mycobacterium tuberculosis RNA Expression Patterns in Sputum Bacteria Indicate Secreted Esx Factors Contributing to Growth are Highly Expressed in Active Disease

    PubMed Central

    Bukka, Archana; Price, Christopher T. D.; Kernodle, Douglas S.; Graham, James E.

    2012-01-01

    To identify factors contributing to the ability of tubercle bacilli to grow in the lung during active infection, we analyzed RNA expression patterns in bacteria present in patient sputum. Prominent among bacterial transcripts identified were those encoding secreted peptides of the Esat-6 subfamily that includes EsxK and EsxL (Rv1197 and Rv1198). H37Rv esxKL and esxJI transcripts were differentially expressed under different growth conditions, and disruption of these genes altered growth phase kinetics in typical laboratory batch broth cultures. These growth defects, including the reduced intracellular growth of an ΔesxKL mutant in primary human macrophages, were reversed by either low multiplicity co-infection or co-culture with wild-type bacteria, demonstrating the ability of the secreted factors to rescue isogenic mutants. Complementing either only esxL or esxI alone (Rv1198 or Rv1037c) also reduced observed growth defects, indicating these genes encode factors capable of contributing to growth. Our studies indicate that the Mycobacterium tuberculosis Mtb9.9 family secreted factors EsxL and EsxI can act in trans to modulate growth of intracellular bacteria, and are highly expressed during active human lung infection. PMID:22291682

  19. Sex-related trends in non-conversion of new smear-positive tuberculosis patients in the Free State, South Africa.

    PubMed

    Heunis, J C; Kigozi, N G; van der Merwe, S; Chikobvu, P; Beyers, N

    2014-03-21

    Contexte : Province de l'Etat Libre, Afrique du Sud.Objectif : Examiner les tendances en fonction du sexe de la nonconversion des frottis de crachats après 2 mois chez des nouveaux cas de tuberculose (TB) à frottis positifs pendant une période où la stratégie DOTS opérait.Schéma : Réalisation d'une étude rétrospective de cohorte des cas de TB enregistrés entre 2003 et 2009. La non-conversion était définie par un résultat de frottis positif après 2 mois de traitement. Des analyses descriptives et de modèles linéaires généralisés ont été réalisées et les tendances de non conversion à 2 mois en fonction du sexe ont été estimées.Résultats : Le taux d'ensemble de non conversion était de 12,5% chez les hommes et de 9,3% chez les femmes. La non conversion était significativement associée à l'âge chez les hommes (P < 0,001). Le taux de non conversion a significativement diminué entre 2003 et 2009 de 15,9% à 10,8% chez les hommes (P < 0,001) et de 12% à 6,6% chez les femmes (P < 0,001). Le taux moyen de déclin de la non-conversion était plus élevé chez les femmes à 1% (IC95% 0,8–1,2%) que chez les hommes à 0,8% (IC95% 0,5–1%). En 2009, le risque de non conversion était plus élevé de 60% chez les hommes (RR 1,60; IC95% 1,37–1,86).Conclusion : Le déclin de la tendance à la non-conversion du frottis de crachats après 2 mois de traitement a mis en évidence le succès relatif de la stratégie DOTS dans la lutte contre la TB, avec un meilleur résultat chez les femmes que chez les hommes. Les interventions devraient tenir compte du sexe et de l'âge des patients afin d'améliorer le taux de conversion du frottis de crachats à 2 mois.

  20. Determinants of consultation, diagnosis and treatment delays among new smear-positive pulmonary tuberculosis patients in Morocco: a cross-sectional study.

    PubMed

    Akrim, M; Bennani, K; Essolbi, A; Sghiar, M; Likos, A; Benmamoun, A; Menzhi, O El; Maaroufi, A

    2014-12-17

    We conducted a cross-sectional survey in 2012 in 12 selected provinces and prefectures in Morocco to determine consultation delay (patient delay), diagnosis delay and treatment delay (health system delays), and factors relating to these delays. The sample included 250 eligible and consenting newly diagnosed smearpositive pulmonary tuberculosis patients who were interviewed at the time of their registration within Diagnosis of Tuberculosis and Respiratory Diseases Reference Centers (CDTMR) or Integrated Health Centers (CSI) using a pretested and structured questionnaire. The median total delay was 46 days [inter-quartile interval (IQI) = 29-84 days]. Patient delay (median = 20; IQI = 8-47 days) was higher than health system delay (median=15; IIQ = 7-35 days). Being illiterate, thinking symptoms will disappear by themselves; having financial constraints and feeling fear of diagnosis or social isolation were associated with patient delay. Consulting first in the private sector or having 3 or more consultations before diagnosis was associated with health system delay.

  1. Evaluation of Xpert® MTB/RIF Assay in Induced Sputum and Gastric Lavage Samples from Young Children with Suspected Tuberculosis from the MVA85A TB Vaccine Trial

    PubMed Central

    Geldenhuys, Hennie; Schmidt, Bey-Marrie; Luabeya, Angelique Kany Kany; Mulenga, Humphrey; Scriba, Thomas J.; Hanekom, Willem A.; Mahomed, Hassan; McShane, Helen; Hatherill, Mark

    2015-01-01

    Objective Diagnosis of childhood tuberculosis is limited by the paucibacillary respiratory samples obtained from young children with pulmonary disease. We aimed to compare accuracy of the Xpert® MTB/RIF assay, an automated nucleic acid amplification test, between induced sputum and gastric lavage samples from young children in a tuberculosis endemic setting. Methods We analyzed standardized diagnostic data from HIV negative children younger than four years of age who were investigated for tuberculosis disease near Cape Town, South Africa [2009–2012]. Two paired, consecutive induced sputa and early morning gastric lavage samples were obtained from children with suspected tuberculosis. Samples underwent Mycobacterial Growth Indicator Tube [MGIT] culture and Xpert MTB/RIF assay. We compared diagnostic yield across samples using the two-sample test of proportions and McNemar’s χ2 test; and Wilson’s score method to calculate sensitivity and specificity. Results 1,020 children were evaluated for tuberculosis during 1,214 admission episodes. Not all children had 4 samples collected. 57 of 4,463[1.3%] and 26 of 4,606[0.6%] samples tested positive for Mycobacterium tuberculosis on MGIT culture and Xpert MTB/RIF assay respectively. 27 of 2,198[1.2%] and 40 of 2,183[1.8%] samples tested positive [on either Xpert MTB/RIF assay or MGIT culture] on induced sputum and gastric lavage samples, respectively. 19/1,028[1.8%] and 33/1,017[3.2%] admission episodes yielded a positive MGIT culture or Xpert MTB/RIF assay from induced sputum and gastric lavage, respectively. Sensitivity of Xpert MTB/RIF assay was 8/30[26.7%; 95% CI: 14.2–44.4] for two induced sputum samples and 7/31[22.6%; 11.4–39.8] [p = 0.711] for two gastric lavage samples. Corresponding specificity was 893/893[100%;99.6–100] and 885/890[99.4%;98.7–99.8] respectively [p = 0.025]. Conclusion Sensitivity of Xpert MTB/RIF assay was low, compared to MGIT culture, but diagnostic performance of Xpert MTB

  2. Benralizumab for chronic obstructive pulmonary disease and sputum eosinophilia: a randomised, double-blind, placebo-controlled, phase 2a study

    PubMed Central

    Brightling, Christopher E; Bleecker, Eugene R; Panettieri, Reynold A; Bafadhel, Mona; She, Dewei; Ward, Christine K; Xu, Xiao; Birrell, Claire; van der Merwe, René

    2016-01-01

    Summary Background Chronic obstructive pulmonary disease (COPD) is associated with eosinophilic airway inflammation in 10–20% of patients. Benralizumab, an anti-interleukin-5 receptor α monoclonal antibody, depletes blood and sputum eosinophils. We aimed to establish whether benralizumab reduces acute exacerbations of COPD in patients with eosinophilia and COPD. Methods We did this randomised, double-blind, placebo-controlled, phase 2a study between Nov 18, 2010, and July 13, 2013, at 26 sites in the UK, Poland, Germany, Canada, the USA, Denmark, and Spain. Adults aged 40–85 years, with moderate-to-severe COPD, at least one acute exacerbation of COPD, and a sputum eosinophil count of 3·0% or more within the previous year, were randomly assigned (1:1) via computer-generated permuted block randomisation (block size of four), with an interactive voice or web-response system, to receive placebo or 100 mg benralizumab subcutaneously, every 4 weeks (three doses), then every 8 weeks (five doses) over 48 weeks. Study site personnel included in study assessments, participants, and data analysts, were masked to treatment allocation. The primary endpoint was the annualised rate of acute exacerbations of COPD at week 56, defined as the number of acute exacerbations divided by total duration of person-year follow-up. Secondary and exploratory endpoints included COPD-specific Saint George’s Respiratory Questionnaire (SGRQ-C), Chronic Respiratory Questionnaire self-administered standardised format (CRQ-SAS), pre-bronchodilator forced expiratory volume in 1 second (FEV1), and safety. We did a prespecified subgroup analysis by baseline blood eosinophil count. Analyses were by intention to treat and per-protocol. This trial is registered with ClinicalTrials.gov, number NCT01227278. Findings We randomly assigned 101 patients to receive placebo (n=50) or benralizumab (n=51), of whom 88 (87%) patients completed the study. Six patients who completed the study were excluded from

  3. Added value of molecular assay Xpert MTB/RIF compared to sputum smear microscopy to assess the risk of tuberculosis transmission in a low-prevalence country.

    PubMed

    Opota, O; Senn, L; Prod'hom, G; Mazza-Stalder, J; Tissot, F; Greub, G; Jaton, K

    2016-07-01

    Airborne precautions are required at hospital admission for patients with suspected pulmonary tuberculosis. The isolation is maintained until 3 serially collected sputum smears are acid-fast bacilli negative, a time- and labor-intensive method with limited sensitivity and specificity, which has a great impact on patient flow management. We evaluated the possibility of replacing the result of microscopy by the semiquantitative result of the molecular point-of-care test Xpert MTB/RIF to assess patients' transmission risk to quickly guide airborne isolation decisions in low-endemic countries. The performance of the Xpert MTB/RIF, used as a first-line test, was compared to the results of microscopy for specimens (n=242) collected from May 2010 to December 2014 in Lausanne, Switzerland. The sensitivity and specificity of Xpert MTB/RIF were 91.5% (65/71) and 99.6% (170/171), respectively, vs. 64.8% (46/71) and 94.2% (161/171) for microscopy. Samples with negative Xpert MTB/RIF were all smear negative for Mycobacterium tuberculosis (negative predictive value, 100%). The semiquantitative results of Xpert MTB/RIF-high, medium, low or very low-were found to correlate with acid-fast bacilli detection: positive predictive value of 100% (6/6), 96.5% (27/28), 52.2% (12/23) and 11.1% (1/9) respectively. Finally, when including clinical criteria, we identified 11 smear-negative but Xpert MTB/RIF-positive patients with a significant transmission potential. In conclusion, our data support the introduction of an Xpert MTB/RIF-based strategy as a replacement of smear microscopy for a faster and more accurate management of tuberculosis patients' transmission risk in a low-prevalence country. PMID:27139592

  4. Added value of molecular assay Xpert MTB/RIF compared to sputum smear microscopy to assess the risk of tuberculosis transmission in a low-prevalence country.

    PubMed

    Opota, O; Senn, L; Prod'hom, G; Mazza-Stalder, J; Tissot, F; Greub, G; Jaton, K

    2016-07-01

    Airborne precautions are required at hospital admission for patients with suspected pulmonary tuberculosis. The isolation is maintained until 3 serially collected sputum smears are acid-fast bacilli negative, a time- and labor-intensive method with limited sensitivity and specificity, which has a great impact on patient flow management. We evaluated the possibility of replacing the result of microscopy by the semiquantitative result of the molecular point-of-care test Xpert MTB/RIF to assess patients' transmission risk to quickly guide airborne isolation decisions in low-endemic countries. The performance of the Xpert MTB/RIF, used as a first-line test, was compared to the results of microscopy for specimens (n=242) collected from May 2010 to December 2014 in Lausanne, Switzerland. The sensitivity and specificity of Xpert MTB/RIF were 91.5% (65/71) and 99.6% (170/171), respectively, vs. 64.8% (46/71) and 94.2% (161/171) for microscopy. Samples with negative Xpert MTB/RIF were all smear negative for Mycobacterium tuberculosis (negative predictive value, 100%). The semiquantitative results of Xpert MTB/RIF-high, medium, low or very low-were found to correlate with acid-fast bacilli detection: positive predictive value of 100% (6/6), 96.5% (27/28), 52.2% (12/23) and 11.1% (1/9) respectively. Finally, when including clinical criteria, we identified 11 smear-negative but Xpert MTB/RIF-positive patients with a significant transmission potential. In conclusion, our data support the introduction of an Xpert MTB/RIF-based strategy as a replacement of smear microscopy for a faster and more accurate management of tuberculosis patients' transmission risk in a low-prevalence country.

  5. Mycological Profile of Sputum of HIV Positive Patients with Lower Respiratory Tract Infection and its Correlation with CD4+ T Lymphocyte Count

    PubMed Central

    Chandwani, Jyotsna; Vyas, Nitya; Hooja, Saroj; Maheshwari, Rakesh

    2016-01-01

    Introduction Fungal respiratory infections are important cause of mortality and morbidity among HIV positive individuals. They account for up to 70% of illness in Acquired Immunodeficiency Disease Syndrome cases (AIDS). The range of illness varies from asymptomatic mucosal candidiasis to overwhelming disseminated infections. In these patients dissemination of fungus leads to very serious outcomes hence, it is important to have the knowledge of prevailing profile of fungus causing infections, so that it can be treated at the onset. Low CD4+ T lymphocyte count is an excellent indicator of decreased immunity and can also be helpful to predict opportunistic fungal respiratory infections and other complications. Aim To define the fungal aetiology of lower respiratory tract infections in HIV positive patients and to correlate the occurrence of different fungi with CD4+ T lymphocyte count. Materials and Methods This was a cross sectional study conducted between May 2014 to April 2015, on 180 treatment naive HIV seropositive patients with lower respiratory tract infections attending the Integrated Counselling and Testing Centre, SMS Medical College, Jaipur, Rajasthan. Early morning expectorated and induced sputum samples were collected and processed for isolation and identification of fungal species. CD4+ T lymphocyte count estimation was done by BD FACS Calibur. Results Fungal species were isolated from 155 (86.1%) patients. The most common isolate was Candida albicans (31.7%), followed by Aspergillus niger (17.7%) and Aspergillus flavus (10%). The fungal species were most commonly isolated from patients with CD4+ T lymphocyte cell less than 200 cells/μl. Conclusion Fungal infections were seen in 86.1% of HIV positive patients with lower respiratory tract infections hence, high level of clinical suspicion for fungal aetiology of respiratory infections in HIV positive patients should be kept in mind. PMID:27790435

  6. Intensified tuberculosis case finding amongst vulnerable communities in southern India.

    PubMed

    Reddy, K K; Ananthakrishnan, R; Jacob, A G; Das, M; Isaakidis, P; Kumar, A M V

    2015-12-21

    India mainly uses passive case finding to detect tuberculosis (TB) patients through the Revised National Tuberculosis Control Programme (RNTCP). An intensified case finding (ICF) intervention was conducted among vulnerable communities in two districts of Karnataka during July-December 2013; 658 sputum smear-positive TB cases were detected. The number of smear-positive cases detected increased by 8.8% relative to the pre-intervention period (July-December 2012) in intervention communities as compared to an 8.6% decrease in communities without the ICF intervention. ICF activities brought TB services closer to vulnerable communities, moderately increasing TB case detection rates.

  7. ENHANCED REMEDIATION DEMONSTRATIONS AT HILL AFB: INTRODUCTION

    EPA Science Inventory

    Nine enhanced aquifer remediation technologies were demonstrated side-by-side at a Hill Air Force Base Chemical Disposal Pit/Fire Training Area site. The demonstrations were performed inside 3 x 5 m cells isolated from the surrounding shallow aquifer by steel piling. The site w...

  8. Orbiter 101 'Enterprise' lands at Edwards AFB

    NASA Technical Reports Server (NTRS)

    1977-01-01

    The rear wheels of the Space Shuttle Orbiter 101 raise desert sand on the landing strip at Edwards Air Force Base as the 'Enterprise' completes the fourth of five scheduled Approach and Landing Test (ALT) free flights. This mission marked the first occasion of the orbiter to fly minus tail cone. The unpowered phase, with Astronauts Joe H. Engle, commander, and Richard H. Truly, pilot, controlling the Enterprise, took two-minutes and 34 seconds. One of the T-38 chase planes that remained with the Shuttle craft for the mission's duration is partially visible at right.

  9. X-31 in Flight over Edwards AFB

    NASA Technical Reports Server (NTRS)

    1994-01-01

    One of two X-31 Enhanced Fighter Maneuverability Demonstrator aircraft, flown by an international test organization at NASA's Dryden Flight Research Center, Edwards, California, turns tightly over the desert floor on a research flight. The aircraft obtained data that may apply to the design and development of highly-maneuverable aircraft of the future. The X-31 had a three-axis thrust-vectoring system, coupled with advanced flight controls, to allow it to maneuver tightly at very high angles of attack. The X-31 Enhanced Fighter Maneuverability (EFM) demonstrator flew at the Ames- Dryden Flight Research Facility, Edwards, California (redesignated the Dryden Flight Research Center in 1994) from February 1992 until 1995 and before that at the Air Force's Plant 42 in Palmdale, California. The goal of the project was to provide design information for the next generation of highly maneuverable fighter aircraft. This program demonstrated the value of using thrust vectoring (directing engine exhaust flow) coupled with an advanced flight control system to provide controlled flight to very high angles of attack. The result was a significant advantage over most conventional fighters in close-in combat situations. The X-31 flight program focused on agile flight within the post-stall regime, producing technical data to give aircraft designers a better understanding of aerodynamics, effectiveness of flight controls and thrust vectoring, and airflow phenomena at high angles of attack. Stall is a condition of an airplane or an airfoil in which lift decreases and drag increases due to the separation of airflow. Thrust vectoring compensates for the loss of control through normal aerodynamic surfaces that occurs during a stall. Post-stall refers to flying beyond the normal stall angle of attack, which in the X-31 was at a 30-degree angle of attack. During Dryden flight testing, the X-31 aircraft established several milestones. On November 6, 1992, the X-31 achieved controlled flight at a 70-degree angle of attack. On April 29, 1993, the second X-31 successfully executed a rapid minimum-radius, 180-degree turn using a post-stall maneuver, flying well beyond the aerodynamic limits of any conventional aircraft. This revolutionary maneuver has been called the 'Herbst Maneuver' after Wolfgang Herbst, a German proponent of using post-stall flight in air-to-air combat. It is also called a 'J Turn' when flown to an arbitrary heading change. The aircraft was flown in tactical maneuvers against an F/A-18 and other tactical aircraft as part of the test flight program. During November and December 1993, the X-31 reached a supersonic speed of Mach 1.28. In 1994, the X-31 program installed software to demonstrate quasi-tailless operation. The X-31 flight test program was conducted by an international test organization (ITO) managed by the Advanced Research Projects Office (ARPA), known as the Defense Advanced Research Projects Office (DARPA) before March 1993. The ITO included the U.S. Navy and U.S. Air Force, Rockwell Aerospace, the Federal Republic of Germany, Daimler-Benz (formerly Messerschmitt-Bolkow-Blohm and Deutsche Aerospace), and NASA. Gary Trippensee was the ITO director and NASA Project Manager. Pilots came from participating organizations. The X-31 was 43.33 feet long with a wingspan of 23.83 feet. It was powered by a single General Electric P404-GE-400 turbofan engine that produced 16,000 pounds of thrust in afterburner.

  10. Efficacy and safety of letosteine in the treatment of sputum thickening and expectoration difficulty in patients with respiratory diseases: a multicenter, randomized, double-masked, double dummy, positive drug parallel controlled trial.

    PubMed

    Zhou, Yan; Sun, Shenghua; Liu, Hua; Cui, Liying; Chang, Xiaoyue; Sun, Zhenqiu

    2014-11-01

    This study aimed to compare the efficacy and safety of letosteine and ambroxol hydrochloride for the treatment of sputum thickening and expectoration difficulty due to either acute or chronic respiratory diseases. Patients (n = 240) were randomized to receive either letosteine + placebo (50 mg thrice daily, Group A) or ambroxol hydrochloride + placebo (30 mg thrice daily, Group B) orally for 5-14 days. The primary outcomes comprised the total effectiveness rate and the total improvement rate. Secondary outcomes included: post-treatment IgA level changes and post-treatment therapeutic evaluation scoring of clinical symptoms. The full analysis set (FAS) comprised 113 patients in Group A and 116 in Group B. The total effectiveness rates were 95.58% for Group A and 95.69% for Group B. The total improvement rates were 99.12% and 99.14% for Group A and Group B, respectively. There were no significant differences between the two groups for any of the primary or secondary outcomes in either the FAS or the per protocol populations (PPS; all P-values > 0.05). Letosteine and ambroxol hydrochloride provided equivalent efficacy and safety in the treatment of sputum thickening and expectoration difficulty due to either acute or chronic respiratory diseases.

  11. Efficacy and safety of letosteine in the treatment of sputum thickening and expectoration difficulty in patients with respiratory diseases: a multicenter, randomized, double-masked, double dummy, positive drug parallel controlled trial.

    PubMed

    Zhou, Yan; Sun, Shenghua; Liu, Hua; Cui, Liying; Chang, Xiaoyue; Sun, Zhenqiu

    2014-11-01

    This study aimed to compare the efficacy and safety of letosteine and ambroxol hydrochloride for the treatment of sputum thickening and expectoration difficulty due to either acute or chronic respiratory diseases. Patients (n = 240) were randomized to receive either letosteine + placebo (50 mg thrice daily, Group A) or ambroxol hydrochloride + placebo (30 mg thrice daily, Group B) orally for 5-14 days. The primary outcomes comprised the total effectiveness rate and the total improvement rate. Secondary outcomes included: post-treatment IgA level changes and post-treatment therapeutic evaluation scoring of clinical symptoms. The full analysis set (FAS) comprised 113 patients in Group A and 116 in Group B. The total effectiveness rates were 95.58% for Group A and 95.69% for Group B. The total improvement rates were 99.12% and 99.14% for Group A and Group B, respectively. There were no significant differences between the two groups for any of the primary or secondary outcomes in either the FAS or the per protocol populations (PPS; all P-values > 0.05). Letosteine and ambroxol hydrochloride provided equivalent efficacy and safety in the treatment of sputum thickening and expectoration difficulty due to either acute or chronic respiratory diseases. PMID:25985582

  12. Notification Rate of Tuberculosis among Migrants in China 2005–2014: A Systematic Review and Meta-analysis

    PubMed Central

    Sun, Yi-Xuan; Zhu, Lei; Lu, Zu-Hong; Jia, Zhong-Wei

    2016-01-01

    Background: Migrations have been reported to be associated with the high risk of tuberculosis (TB), but there is no systematic analysis of the available data for TB among migrant in China. The aim of this study was to examine the notification rate of active and sputum smear-positive TB by a systematic review and meta-analysis. Methods: A systematic review and meta-analysis were performed to examine the notification rate of active and sputum smear-positive TB among migrants in China. Two reviewers searched the cross-sectional studies published in PubMed, EMBASE, SciFinder, and Web of Science in English and in CNKI and Wanfang databases in Chinese. Pooled estimates of notification rate of TB among migrants were calculated using a random effects model. Meta-regression analysis and subgroup analysis stratified by year, region were also performed. Results: Seventy eligible studies met the inclusion criteria for the final analysis. The overall notification rate of active TB and sputum smear-positive cases among migrants were 53.12 (95% confidence interval [CI]: 47.32–59.63) and 24.53 (95% CI: 22.01–27.34) per 100,000 populations, respectively. The notification rate of active TB significantly increased from 50.95 (95% CI: 41.11–63.14) per 100,000 populations in 2005 to 84.62 (95% CI: 78.00–91.80) per 100,000 populations in 2014 while that of smear-positive TB was constant during the study time (P = 0.79). The geographic difference was identified both for active and sputum smear-positive TB, with the higher notification rates mainly distributing along the eastern coastal areas. Conclusions: The pooled estimate of active TB and sputum smear-positive TB among migrants was lower than the national notification rate among general population, but the gap between our data and national notification rate among general population is narrowed down during 2005–2014. PMID:27453237

  13. Experience of active tuberculosis case finding in nearly 5 million households in India.

    PubMed

    Prasad, B M; Satyanarayana, S; Chadha, S S; Das, A; Thapa, B; Mohanty, S; Pandurangan, S; Babu, E R; Tonsing, J; Sachdeva, K S

    2016-03-21

    In India, to increase tuberculosis (TB) case detection under the National Tuberculosis Programme, active case finding (ACF) was implemented by the Global Fund-supported Project Axshya, among high-risk groups in 300 districts. Between April 2013 and December 2014, 4.9 million households covering ~20 million people were visited. Of 350 047 presumptive pulmonary TB cases (cough of ⩾2 weeks) identified, 187 586 (54%) underwent sputum smear examination and 14 447 (8%) were found to be smear-positive. ACF resulted in the detection of a large number of persons with presumptive pulmonary TB and smear-positive TB. Ensuring sputum examination of all those with presumptive TB was a major challenge.

  14. Detection of Mycobacterium tuberculosis (MTB) in Fecal Specimens From Adults Diagnosed With Pulmonary Tuberculosis Using the Xpert MTB/Rifampicin Test

    PubMed Central

    Kokuto, Hiroyuki; Sasaki, Yuka; Yoshimatsu, Shoji; Mizuno, Kazue; Yi, Lina; Mitarai, Satoshi

    2015-01-01

    Background. The Xpert Mycobacterium tuberculosis (MTB)/rifampicin (RIF) is a fully automated diagnostic test that allows for the detection of MTB including its RIF resistance. Although the test is used for the diagnosis of tuberculosis (TB) in sputum samples worldwide, studies using fecal specimens are scarce. We therefore evaluated the efficacy of the Xpert MTB/RIF test for detection of MTB in fecal specimens obtained from adult pulmonary TB patients, confirmed by culture and/or molecular diagnostic methods. Methods. We conducted a retrospective case-control study to provide proof-of-concept regarding the efficacy of the Xpert MTB/RIF test using fecal samples for diagnosing pulmonary TB via detection of MTB in adult patients (≥20 years) at the Fukujuji Hospital in Tokyo, Japan. Results. Fecal specimens were obtained from 56 active pulmonary TB patients (including 48 sputum smear-positive and 8 sputum smear-negative patients), 10 non-TB patients (including 4 Myocobacterium avium complex infections), and 27 healthy individuals who were exposed to active pulmonary TB patients. The sensitivity of the fecal Xpert MTB/RIF was 100% (81.7%–100%) for detection of MTB in specimens from sputum smear-positive (1+ to 3+) patients, 81.0% (58.1%–94.6%) in specimens from sputum smear scanty positive patients, and 50.0% (15.7%–84.3%) in specimens from sputum smear-negative patients. Meanwhile, each of the fecal specimens from the non-TB group was negative for MTB (specificity 100%; 95% confidence interval, 86.2–100). Conclusions. The fecal Xpert MTB/RIF test could detect MTB in a large proportion of smear-positive pulmonary TB patients, without frequent false-positive results at a TB referral hospital in Japan. PMID:26125035

  15. The effect of HIV and antiretroviral therapy on characteristics of pulmonary tuberculosis in northern Malawi: a cross-sectional study

    PubMed Central

    2014-01-01

    Background HIV infection reduces the likelihood that individuals with pulmonary tuberculosis are smear positive and that they have cavitatory disease. Antiretroviral therapy (ART) may shift the pattern of disease to be more similar to that of HIV negative patients. This would aid diagnosis- which often depends on sputum smears – but would also increase infectiousness. We assessed the effect of HIV and ART on smear positivity and cavitatory disease in laboratory-confirmed pulmonary TB patients. Methods Three sputum samples were collected per pulmonary TB suspect and were examined using microscopy and culture. Chest radiographs were available for a subset of patients as part of another study. The effect of HIV and ART status on sputum smear positivity and lung cavitation were evaluated using multivariable logistic regression. Results Of 1024 laboratory-confirmed pulmonary TB patients who were identified between January 2005 and December 2011, 766 had HIV and ART status available. Positive sputum smears were significantly more common among HIV negative individuals than HIV positive individuals (adjusted OR = 2.91, 95% CI 1.53 – 5.55). Compared to those HIV positive but not on ART, patients on ART were more likely to be smear positive (adjusted OR = 2.33, 95% CI 1.01 – 5.39) if they had been on ART ≤ 6 months, but only slightly more likely to be smear positive (adjusted OR = 1.43, 95% CI 0.68 – 2.99) if they were on ART > 6 months. HIV negative patients were more likely than HIV positive patients to have cavitatory disease (adjusted OR = 1.97, 95% CI 1.20 – 3.23). Patients on ART > 6 months had a slight increase in cavitatory disease compared to HIV positive patients not on ART (adjusted OR = 1.68, CI 0.78 – 3.63). Conclusions HIV infection is associated with less smear positivity and cavitation in pulmonary TB patients. Among HIV positive patients, the use of ART shifts the presentation of disease towards that seen in HIV

  16. Peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) assay for specific detection of Mycobacterium immunogenum and DNA-FISH assay for analysis of pseudomonads in metalworking fluids and sputum.

    PubMed

    Selvaraju, Suresh B; Kapoor, Renuka; Yadav, Jagjit S

    2008-01-01

    Specific and rapid detection and quantification of mycobacteria in contaminated metalworking fluid (MWF) are problematic due to complexity of the matrix and heavy background co-occurring microflora. Furthermore, cross-reactivity among neighboring species of Mycobacterium makes species differentiation difficult for this genus. Here, we report for the first time a species-specific peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) method for Mycobacterium immunogenum, a non-tuberculous Mycobacterium species prevalent in MWF and implicated in occupational lung disease hypersensitivity pneumonitis and pseudo-outbreaks. A novel species-specific 14-bp PNA probe was designed for M. immunogenum based on its 16S rRNA gene sequence and was validated for specificity, by testing against a panel of other phylogenetically closely related rapidly growing mycobacteria and representative species of gram-positive, gram-negative, and acid fast organisms. In addition, a DNA-FISH protocol was optimized for co-detection of Pseudomonas, the most predominantly co-occurring genus in contaminated MWF. Reliable quantification for both the test organisms was achieved at or above a cell density of 10(3)cellsml(-1), a recognized minimum limit for microscopic quantification. The mycobacterial PNA-FISH assay was successfully adapted to human sputum demonstrating its potential for clinical diagnostic applications in addition to industrial MWF monitoring, to assess MWF-associated exposures and pseudo-outbreaks.

  17. Rapid and simultaneous detection of Mycobacterium tuberculosis complex and Beijing/W genotype in sputum by an optimized DNA extraction protocol and a novel multiplex real-time PCR.

    PubMed

    Leung, Eric T Y; Zheng, L; Wong, Rity Y K; Chan, Edward W C; Au, T K; Chan, Raphael C Y; Lui, Grace; Lee, Nelson; Ip, Margaret

    2011-07-01

    Rapid diagnosis and genotyping of Mycobacterium tuberculosis by molecular methods are often limited by the amount and purity of DNA extracted from body fluids. In this study, we evaluated 12 DNA extraction methods and developed a highly sensitive protocol for mycobacterial DNA extraction directly from sputa using surface-coated magnetic particles. We have also developed a novel multiplex real-time PCR for simultaneous identification of M. tuberculosis complex and the Beijing/W genotype (a hypervirulent sublineage of M. tuberculosis) by using multiple fluorogenic probes targeting both the M. tuberculosis IS6110 and the Rv0927c-pstS3 intergenic region. With reference strains and clinical isolates, our real-time PCR accurately identified 20 non-Beijing/W and 20 Beijing/W M. tuberculosis strains from 17 different species of nontuberculosis Mycobacterium (NTM). Further assessment of our DNA extraction protocol and real-time PCR with 335 nonduplicate sputum specimens correctly identified all 74 M. tuberculosis culture-positive specimens. In addition, 15 culture-negative specimens from patients with confirmed tuberculosis were also identified. No cross-reactivity was detected with NTM specimens (n = 31). The detection limit of the assay is 10 M. tuberculosis bacilli, as determined by endpoint dilution analysis. In conclusion, an optimized DNA extraction protocol coupled with a novel multiprobe multiplex real-time PCR for the direct detection of M. tuberculosis, including Beijing/W M. tuberculosis, was found to confer high sensitivity and specificity. The combined procedure has the potential to compensate for the drawbacks of conventional mycobacterial culture in routine clinical laboratory setting, such as the lengthy incubation period and the limitation to viable organisms.

  18. An approach to the problems of diagnosing and treating adult smear-negative pulmonary tuberculosis in high-HIV-prevalence settings in sub-Saharan Africa.

    PubMed Central

    Harries, A. D.; Maher, D.; Nunn, P.

    1998-01-01

    The overlap between the populations in sub-Saharan Africa infected with human immunodeficiency virus (HIV) and Mycobacterium tuberculosis has led to an upsurge in tuberculosis cases over the last 10 years. The relative increase in the proportion of notified sputum-smear-negative pulmonary tuberculosis (PTB) cases is greater than that of sputum-smear-positive PTB cases. This is a consequence of the following: the association between decreased host immunity and reduced sputum smear positivity; the difficulty in excluding other HIV-related diseases when making the diagnosis of smear-negative PTB; and an increase in false-negative sputum smears because of overstretched resources. This article examines problems in the diagnosis and treatment of smear-negative PTB in high-HIV-prevalence areas in sub-Saharan Africa. The main issues in diagnosis include: the criteria used to diagnose smear-negative PTB; the degree to which clinicians actually follow these criteria in practice; and the problem of how to exclude other respiratory diseases that can resemble, and be misdiagnosed as, smear-negative PTB. The most important aspect of the treatment of smear-negative PTB patients is abandoning 12-month "standard" treatment regimens in favour of short-course chemotherapy. Operational research is necessary to determine the most cost-effective approaches to the diagnosis and treatment of smear-negative PTB. Nevertheless, substantial improvement could be obtained by implementing the effective measures already available, such as improved adherence to diagnostic and treatment guidelines. PMID:10191561

  19. Implementing intensified tuberculosis case-finding among street-connected youth and young adults in Kenya

    PubMed Central

    Mercer, T.; Kimani, S.; Braitstein, P.; Buziba, N.; Carter, E. J.

    2016-01-01

    Background: Street-connected youth and young adults (SCY) suffer a myriad of health problems. In Kenya, SCY are at high risk for tuberculosis (TB) due to their congregate living situations. TB screening is not routinely implemented in SCY and there has been no published literature on the burden of TB in SCY in western Kenya. Program description: In 2011, the AMPATH TB Program, an experienced TB screening program, partnered with the Tumaini Center, a trusted street youth organization, to conduct intensified case finding (ICF) for pulmonary TB among SCY. Our program aimed to investigate the numbers of SCY who reported symptoms and those diagnosed with smear-positive pulmonary TB, and link SCY with TB to treatment. Results: Of 116 SCY who were screened, 114 (98%) had a positive questionnaire; 104 (90%) provided a spot sputum sample, 39 (34%) provided a morning sputum sample, and 111 (97%) reported cough of >2 weeks. One street youth tested smear-positive for TB and was treated through to cure. Conclusions: Implementing TB ICF is feasible in low-resource settings through unique collaborations between health care programs and community-based organizations. In addition to identifying smear-positive TB, our program uncovered a high burden of respiratory symptoms among SCY in Eldoret, Kenya. PMID:27358809

  20. Cystic Fibrosis (CF) Respiratory Screen: Sputum

    MedlinePlus

    ... Cystic Fibrosis (CF) Chloride Sweat Test Lungs and Respiratory System Cystic Fibrosis: Diet and Nutrition Cystic Fibrosis Cystic Fibrosis: Diet and Nutrition Lungs and Respiratory System Contact Us Print Resources Send to a friend ...

  1. Evaluation of the Speed-oligo Direct Mycobacterium tuberculosis Assay for Molecular Detection of Mycobacteria in Clinical Respiratory Specimens

    PubMed Central

    Lara-Oya, Ana; Mendoza-Lopez, Pablo; Rodriguez-Granger, Javier; Fernández-Sánchez, Ana María; Bermúdez-Ruiz, María Pilar; Toro-Peinado, Inmaculada; Palop-Borrás, Begoña; Navarro-Marí, Jose María

    2013-01-01

    We present the first evaluation of a novel molecular assay, the Speed-oligo Direct Mycobacterium tuberculosis (SO-DMT) assay, which is based on PCR combined with a dipstick for the detection of mycobacteria and the specific identification of M. tuberculosis complex (MTC) in respiratory specimens. A blind evaluation was carried out in two stages: first, under experimental conditions on convenience samples comprising 20 negative specimens, 44 smear- and culture-positive respiratory specimens, and 11 sputa inoculated with various mycobacterium-related organisms; and second, in the routine workflow of 566 fresh respiratory specimens (4.9% acid-fast bacillus [AFB] smear positives, 7.6% MTC positives, and 1.8% nontuberculous mycobacteria [NTM] culture positives) from two Mycobacterium laboratories. SO-DMT assay showed no reactivity in any of the mycobacterium-free specimens or in those with mycobacterium-related organisms. Compared to culture, the sensitivity in the selected smear-positive specimens was 0.91 (0.92 for MTC and 0.90 for NTM), and there was no molecular detection of NTM in a tuberculosis case or vice versa. With respect to culture and clinical data, the sensitivity, specificity, and positive and negative predictive values for the SO-DMT system in routine specimens were 0.76 (0.93 in smear positives [1.0 for MTC and 0.5 for NTM] and 0.56 in smear negatives [0.68 for MTC and 0.16 for NTM]), 0.99, 0.85 (1.00 in smear positives and 0.68 in smear negatives), and 0.97, respectively. Molecular misidentification of NTM cases occurred when testing 2 gastric aspirates from two children with clinically but not microbiologically confirmed lung tuberculosis. The SO-DMT assay appears to be a fast and easy alternative for detecting mycobacteria and differentiating MTC from NTM in smear-positive respiratory specimens. PMID:23100355

  2. Detection of lipoarabinomannan as a diagnostic test for tuberculosis.

    PubMed Central

    Sada, E; Aguilar, D; Torres, M; Herrera, T

    1992-01-01

    A coagglutination technique was established for the detection of lipoarabinomannan of Mycobacterium tuberculosis in human serum samples and evaluated for its utility in the diagnosis of tuberculosis at the Instituto Nacional de Enfermedades Respiratorias in Mexico City. The test had a sensitivity of 88% in patients with sputum-smear-positive active pulmonary tuberculosis. The sensitivity in patients with active pulmonary tuberculosis negative for acid-fast bacilli in sputum was 67%. Less favorable results were obtained for patients with AIDS and tuberculosis, with a sensitivity of 57%. The specificity in control patients with lung diseases different from tuberculosis and in healthy subjects was 100%. The positive predictive value was 100%, and the negative predictive value for patients with sputum-positive active pulmonary tuberculosis was 97%. The results of this study suggest that the detection of lipoarabinomannan is an accurate test for the diagnosis of pulmonary tuberculosis. PMID:1401008

  3. Detection of lipoarabinomannan as a diagnostic test for tuberculosis.

    PubMed

    Sada, E; Aguilar, D; Torres, M; Herrera, T

    1992-09-01

    A coagglutination technique was established for the detection of lipoarabinomannan of Mycobacterium tuberculosis in human serum samples and evaluated for its utility in the diagnosis of tuberculosis at the Instituto Nacional de Enfermedades Respiratorias in Mexico City. The test had a sensitivity of 88% in patients with sputum-smear-positive active pulmonary tuberculosis. The sensitivity in patients with active pulmonary tuberculosis negative for acid-fast bacilli in sputum was 67%. Less favorable results were obtained for patients with AIDS and tuberculosis, with a sensitivity of 57%. The specificity in control patients with lung diseases different from tuberculosis and in healthy subjects was 100%. The positive predictive value was 100%, and the negative predictive value for patients with sputum-positive active pulmonary tuberculosis was 97%. The results of this study suggest that the detection of lipoarabinomannan is an accurate test for the diagnosis of pulmonary tuberculosis.

  4. Treatment of multidrug-resistant tuberculosis in China.

    PubMed

    Zhang, L X

    1996-01-01

    During the past decade the number and gravity of tuberculosis (TB) cases has continued to increase, both in developing and industrialized nations. Coupled with the recent emergence of multidrug-resistant tuberculosis (MDR-TB), the possibility that untreatable forms of the disease may become widespread has arisen. In China, the prevalence rate of smear-positive cases from three national surveys in 1979, 1984-1985 and 1990 was 187, 156 and 134/100,000, respectively, thus giving an annual average reduction rate of only 3.0%. This may be due to the accumulation of chronic cases, which is not surprising given that as many as 84.3% of new smear-positive cases received non-organized chemotherapy. To counteract this situation, a strategy was developed in Beijing to practice fully supervised chemotherapy for all new smear-positive cases. This is now 90% with a cure rate also of 90%. As a result, the prevalence rate of smear-positive cases has dropped, with an average annual reduction of 17%. Building upon this success, the World Bank Loan TB Control Project in China has been carried out in 12 provinces with 550 million people since 1992. The main objective of this project is to provide fully supervised, 6-month short-course chemotherapy for all newly detected smear-positive cases. The cure rate based on cohort analysis was 88% in 1993. Complete data are not available on resistance although the initial and acquired resistance rates were 28.1 and 41.1%, respectively. MDR-TB treated with ofloxacin has been increasing since 1992, with 317 cases reported during the period 1992-1995, of which 77% showed sputum conversion.

  5. Tuberculosis case-finding through a village outreach programme in a rural setting in southern Ethiopia: community randomized trial.

    PubMed Central

    Shargie, Estifanos Biru; Mørkve, Odd; Lindtjørn, Bernt

    2006-01-01

    OBJECTIVE: To ascertain whether case-finding through community outreach in a rural setting has an effect on case-notification rate, symptom duration, and treatment outcome of smear-positive tuberculosis (TB). METHODS: We randomly allocated 32 rural communities to intervention or control groups. In intervention communities, health workers from seven health centres held monthly diagnostic outreach clinics at which they obtained sputum samples for sputum microscopy from symptomatic TB suspects. In addition, trained community promoters distributed leaflets and discussed symptoms of TB during house visits and at popular gatherings. Symptomatic individuals were encouraged to visit the outreach team or a nearby health facility. In control communities, cases were detected through passive case-finding among symptomatic suspects reporting to health facilities. Smear-positive TB patients from the intervention and control communities diagnosed during the study period were prospectively enrolled. FINDINGS: In the 1-year study period, 159 and 221 cases of smear-positive TB were detected in the intervention and control groups, respectively. Case-notification rates in all age groups were 124.6/10(5) and 98.1/10(5) person-years, respectively (P = 0.12). The corresponding rates in adults older than 14 years were 207/10(5) and 158/10(5) person-years, respectively (P = 0.09). The proportion of patients with >3 months' symptom duration was 41% in the intervention group compared with 63% in the control group (P<0.001). Pre-treatment symptom duration in the intervention group fell by 55-60% compared with 3-20% in the control group. In the intervention and control groups, 81% and 75%, respectively of patients successfully completed treatment (P = 0.12). CONCLUSION: The intervention was effective in improving the speed but not the extent of case finding for smear-positive TB in this setting. Both groups had comparable treatment outcomes. PMID:16501728

  6. Critique and Summary of the Chanute AFB CBE Project.

    ERIC Educational Resources Information Center

    Himwich, H. A., Ed.

    Chanute Air Force Base was the first military training center to undertake an extensive investigation of the use of the PLATO IV system in technical training. The service test was to compare a conventionally taught course in general vehicle maintenance with a course taught with computer-based instructional materials. Part I of this document is the…

  7. TRANSECT STUDY OF THE INTRINSIC BIOREMEDIATION TEST PLOT: DOVER AFB

    EPA Science Inventory

    The work described in this report is part of a project undertaken by the Bioremediation of Chlorinated Solvents Action Team of the Remediation Technologies Development Forum, a joint U.S. Federal agency-industry collaboration, to study the natural attenuation of chlorinated ethen...

  8. STREAM INJECTION INTO FRACTURE BEDROCK AT LORING AFB

    EPA Science Inventory

    The Quarry at the former Loring Air Force Base in Limestone, Maine, was used for the disposal of drums containing spent chlorinated solvents, mainly tetrachloroethene (PCE). After closure of the base, surface geophysics were used to locate the drums, and they were removed. Subs...

  9. X-31 in Banked Flight over Edwards AFB

    NASA Technical Reports Server (NTRS)

    1994-01-01

    One of two X-31 Enhanced Fighter Maneuverability Demonstrator aircraft, flown by an international test organization at NASA's Dryden Flight Research Center, Edwards, California, turns tightly over the desert floor on a research flight. The aircraft obtained data that may apply to the design and development of highly-maneuverable aircraft of the future. The X-31 had a three-axis thrust-vectoring system, coupled with advanced flight controls, to allow it to maneuver tightly at very high angles of attack. The X-31 Enhanced Fighter Maneuverability (EFM) demonstrator flew at the Ames- Dryden Flight Research Facility, Edwards, California (redesignated the Dryden Flight Research Center in 1994) from February 1992 until 1995 and before that at the Air Force's Plant 42 in Palmdale, California. The goal of the project was to provide design information for the next generation of highly maneuverable fighter aircraft. This program demonstrated the value of using thrust vectoring (directing engine exhaust flow) coupled with an advanced flight control system to provide controlled flight to very high angles of attack. The result was a significant advantage over most conventional fighters in close-in combat situations. The X-31 flight program focused on agile flight within the post-stall regime, producing technical data to give aircraft designers a better understanding of aerodynamics, effectiveness of flight controls and thrust vectoring, and airflow phenomena at high angles of attack. Stall is a condition of an airplane or an airfoil in which lift decreases and drag increases due to the separation of airflow. Thrust vectoring compensates for the loss of control through normal aerodynamic surfaces that occurs during a stall. Post-stall refers to flying beyond the normal stall angle of attack, which in the X-31 was at a 30-degree angle of attack. During Dryden flight testing, the X-31 aircraft established several milestones. On November 6, 1992, the X-31 achieved controlled flight at a 70-degree angle of attack. On April 29, 1993, the second X-31 successfully executed a rapid minimum-radius, 180-degree turn using a post-stall maneuver, flying well beyond the aerodynamic limits of any conventional aircraft. This revolutionary maneuver has been called the 'Herbst Maneuver' after Wolfgang Herbst, a German proponent of using post-stall flight in air-to-air combat. It is also called a 'J Turn' when flown to an arbitrary heading change. The aircraft was flown in tactical maneuvers against an F/A-18 and other tactical aircraft as part of the test flight program. During November and December 1993, the X-31 reached a supersonic speed of Mach 1.28. In 1994, the X-31 program installed software to demonstrate quasi-tailless operation. The X-31 flight test program was conducted by an international test organization (ITO) managed by the Advanced Research Projects Office (ARPA), known as the Defense Advanced Research Projects Office (DARPA) before March 1993. The ITO included the U.S. Navy and U.S. Air Force, Rockwell Aerospace, the Federal Republic of Germany, Daimler-Benz (formerly Messerschmitt-Bolkow-Blohm and Deutsche Aerospace), and NASA. Gary Trippensee was the ITO director and NASA Project Manager. Pilots came from participating organizations. The X-31 was 43.33 feet long with a wingspan of 23.83 feet. It was powered by a single General Electric P404-GE-400 turbofan engine that produced 16,000 pounds of thrust in afterburner.

  10. X-31 #1 in Flight over Edwards AFB

    NASA Technical Reports Server (NTRS)

    1993-01-01

    The first X-31 (Bu. No. 164584) flies over Edwards Air Force Base, California, in 1993. Aircraft 584 completed 292 flights during the Enhanced Fighter Maneuverability (EFM) program before being lost on January 19, 1995 when icing in the nose probe caused the flight control computer to receive bad data. German test pilot Karl-Heinz Lang ejected after the aircraft became uncontrollable. The program continued, using the second aircraft (Bu. No. 164585). The X-31 Enhanced Fighter Maneuverability (EFM) demonstrator flew at the Ames- Dryden Flight Research Facility, Edwards, California (redesignated the Dryden Flight Research Center in 1994) from February 1992 until 1995 and before that at the Air Force's Plant 42 in Palmdale, California. The goal of the project was to provide design information for the next generation of highly maneuverable fighter aircraft. This program demonstrated the value of using thrust vectoring (directing engine exhaust flow) coupled with an advanced flight control system to provide controlled flight to very high angles of attack. The result was a significant advantage over most conventional fighters in close-in combat situations. The X-31 flight program focused on agile flight within the post-stall regime, producing technical data to give aircraft designers a better understanding of aerodynamics, effectiveness of flight controls and thrust vectoring, and airflow phenomena at high angles of attack. Stall is a condition of an airplane or an airfoil in which lift decreases and drag increases due to the separation of airflow. Thrust vectoring compensates for the loss of control through normal aerodynamic surfaces that occurs during a stall. Post-stall refers to flying beyond the normal stall angle of attack, which in the X-31 was at a 30-degree angle of attack. During Dryden flight testing, the X-31 aircraft established several milestones. On November 6, 1992, the X-31 achieved controlled flight at a 70-degree angle of attack. On April 29, 1993, the second X-31 successfully executed a rapid minimum-radius, 180-degree turn using a post-stall maneuver, flying well beyond the aerodynamic limits of any conventional aircraft. This revolutionary maneuver has been called the 'Herbst Maneuver' after Wolfgang Herbst, a German proponent of using post-stall flight in air-to-air combat. It is also called a 'J Turn' when flown to an arbitrary heading change. The aircraft was flown in tactical maneuvers against an F/A-18 and other tactical aircraft as part of the test flight program. During November and December 1993, the X-31 reached a supersonic speed of Mach 1.28. In 1994, the X-31 program installed software to demonstrate quasi-tailless operation. The X-31 flight test program was conducted by an international test organization (ITO) managed by the Advanced Research Projects Office (ARPA), known as the Defense Advanced Research Projects Office (DARPA) before March 1993. The ITO included the U.S. Navy and U.S. Air Force, Rockwell Aerospace, the Federal Republic of Germany, Daimler-Benz (formerly Messerschmitt-Bolkow-Blohm and Deutsche Aerospace), and NASA. Gary Trippensee was the ITO director and NASA Project Manager. Pilots came from participating organizations. The X-31 was 43.33 feet long with a wingspan of 23.83 feet. It was powered by a single General Electric P404-GE-400 turbofan engine that produced 16,000 pounds of thrust in afterburner.

  11. Historical chronology of Hanscom AFB 1941-1986

    SciTech Connect

    Del Papa, E.M.; Warner, M.P.

    1987-11-01

    This report contents are Initial Efforts; Program Guidance; Development, Test and Evaluation (DTandE)/Initial; Operational Test and Evaluation (IOTandE); Production Contract for HAVE A QUICK AN/ARC-164 Modification Kits; Additional HAVE QUICK Requirements; Additional HAVE QUICK Program Guidance; HAVE QUICK Extension; Deployment and Testing; FY 1981 Supplemental Budget Buy of HAVE QUICK Production Units AN/GRC-171 HAVE QUICK Modified Radio Contract; HAVE QUICK for the E-3A AWACS; Program Management Responsibility Transfer (PMRT) for the HAVE QUICK AN/ARC-164 Radio; HAVE QUICK for the Tactical Air Control System (TACS); Additional HAVE QUICK Program Guidance; Air Force Organizational Excellence Award to HAVE QUICK Program Office; HAVE QUICK for the E-3A; HAVE QUICK for NATO; AN/GRC-171 HAVE QUICK Modified Radio; HAVE QUICK Contract F19628-81-c-0151; SEEK BARON; Additional HAVE QUICK Program Guidance; Production; HAVE QUICK Net Rollover Problem; HAVE QUICK System Timing; GRC-171(V)4 Enhancements and Production; HAVE QUICK Technical Order (TO) Issues; HAVE QUICK GRC-171 Multiplexing (MUX) Network; HAVE QUICK for NATO; Navy Problem with HAVE QUICK; Nuclear Hardening, HAVE QUICK ARC-164C; HAVE QUICK GRC-171A(V)2; Midnight Madness; HAVE QUICK GRC-171 Multiplexing (MUX) Network; U.S. Marine Corps HAVE QUICK requirements; HAVE QUICK for NATO; HAVE QUICK Program Costs; and Summary and Conclusion.

  12. Walk-off correction with AFB nonlinear composites

    NASA Astrophysics Data System (ADS)

    Lee, H.-C.; Meissner, H. E.

    2007-04-01

    Beam walk-off in uniaxial and biaxial crystals occurs when the phase normal of the propagating electromagnetic wave deviates from the direction of the Poynting vector. This beam walk-off limits frequency conversion efficiency and restricts the OPO tuning range. The beam walk-off angle in nonlinear single crystals can be alleviated by bonding similar non-linear crystals rotated by 180° with respect to each other. An even number of twisted twins of single crystals is formed that is stress-free and has negligible loss at the AFB® (Adhesive-Free Bond) interfaces. Since no adhesive is employed and the bonding force consists primarily of Van der Waals attractive forces, there is no adverse effect or absorption at the bond interface. The theory of walk-off angles as a function of orientation for uniaxial and biaxial crystals is derived. Correcting beam walk-off by producing an AFB® composite configuration results in more efficient frequency conversion and thereby allows the generation of higher power output of frequency converted radiation for a given input power. Beam correction is demonstrated experimentally for zinc germanium phosphide (ZGP) as representative of a uniaxial nonlinear crystal, and on biaxial KTP crystals. AFB® composites of ZGP with inactive ends of gallium phosphide have been produced in an effort to further improve damage resistance of a ZGP optical parametric oscillator for frequency conversion into the mid-IR range.

  13. NOAA-L satellite arrives at Vandenberg AFB

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Inside the B16-10 spacecraft processing hangar at Vandenberg Air Force Base, Calif., workers oversee the uncrating of the National Oceanic and Atmospheric Administration (NOAA-L) satellite. NOAA-L is part of the Polar-Orbiting Operational Environmental Satellite (POES) program that provides atmospheric measurements of temperature, humidity, ozone and cloud images, tracking weather patterns that affect the global weather and climate. The launch of the NOAA-L satellite is scheduled no earlier than Sept. 12 aboard a Lockheed Martin Titan II rocket. NOAA-L satellite arrives at Vandenberg AFB

    NASA Technical Reports Server (NTRS)

    2000-01-01

    A crated National Oceanic and Atmospheric Administration (NOAA-L) satellite is moved inside the B16-10 spacecraft processing hangar at Vandenberg Air Force Base, Calif. NOAA-L is part of the Polar- Orbiting Operational Environmental Satellite (POES) program that provides atmospheric measurements of temperature, humidity, ozone and cloud images, tracking weather patterns that affect the global weather and climate. The launch of the NOAA-L satellite is scheduled no earlier than Sept. 12 aboard a Lockheed Martin Titan II rocket. NOAA-L satellite arrives at Vandenberg AFB

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Outside the B16-10 spacecraft processing hangar at Vandenberg Air Force Base, Calif., a crated National Oceanic and Atmospheric Administration (NOAA-L) satellite is lowered to the ground before being moved inside. NOAA-L is part of the Polar-Orbiting Operational Environmental Satellite (POES) program that provides atmospheric measurements of temperature, humidity, ozone and cloud images, tracking weather patterns that affect the global weather and climate. The launch of the NOAA-L satellite is scheduled no earlier than Sept. 12 aboard a Lockheed Martin Titan II rocket. NOAA-L satellite arrives at Vandenberg AFB

    NASA Technical Reports Server (NTRS)

    2000-01-01

    A crated National Oceanic and Atmospheric Administration (NOAA-L) satellite arrives at Vandenberg Air Force Base, Calif. It is part of the Polar-Orbiting Operational Environmental Satellite (POES) program that provides atmospheric measurements of temperature, humidity, ozone and cloud images, tracking weather patterns that affect the global weather and climate. The launch of the NOAA-L satellite is scheduled no earlier than Sept. 12 aboard a Lockheed Martin Titan II rocket. AFB (Acid-Fast Bacillus) Smear and Culture

    MedlinePlus

    ... Mycobacteria Smear; Mycobacteria Culture; TB NAAT Formal name: Acid-Fast Bacillus Smear and Culture and Sensitivity; Mycobacteria tuberculosis Nucleic Acid Amplification Test Related tests: TB Screening Tests ; Bacterial ...

  14. AFB's Computerized Travel Aid: Two Years of Research and Development.

    ERIC Educational Resources Information Center

    Uslan, Mark M.; And Others

    1983-01-01

    Progress on the computerized travel aid, an electronic device, using elements of the Polaroid Sonar Camera and a microprocessor, for visually handicapped persons is reviewed, and research on the effectiveness of various models noted. Recommended modifications touch on aspects of mounting, beam shape, and audible outputs. (CL)

  15. Dog Guide Training: AFB Directory of Services Listings

    MedlinePlus

    ... ACB of Minnesota P.O. Box 7341 Minneapolis, MN 55407 mike-hally@hotmail.com http://www.acb. ... New Dimensions, Inc. 100 East 22nd Street Minneapolis, MN 55404-2514 (612) 872-0100 (Local) (612) 872- ...

  16. Tuberculosis situation among tribal population of Car Nicobar, India, 15 years after intensive tuberculosis control project and implementation of a national tuberculosis programme.

    PubMed Central

    Murhekar, M. V.; Kolappan, C.; Gopi, P. G.; Chakraborty, A. K.; Sehgal, S. C.

    2004-01-01

    OBJECTIVE: To assess the tuberculosis (TB) situation in the tribal community of Car Nicobar island 15 years after the national TB programme was implemented in this area after an intensive phase of TB control in 1986. METHODS: The entire population of Car Nicobar was enumerated through a house-to-house survey. Children aged <14 years were tuberculin tested and read for reaction sizes. Individuals aged >15 years were asked about the presence of chest symptoms (cough, chest pain, and unexplained fever for two weeks or longer and haemoptysis), and sputum samples were collected from patients with chest symptoms. Sputum samples were examined for presence of acid-fast bacilli. FINDINGS: Among the 4,543 children enumerated, 4,351 (95.8%) were tuberculin tested and read. Of the 981 children without bacille Calmette-Guerin scars, 161 (16.4%) were infected with TB. A total of 77 cases who were smear-positive for TB were detected from among 10,570 people aged >15 years; the observed smear-positive case prevalence was 728.5 per 100,000. The standardized prevalence of TB infection, annual risk of TB infection, and prevalence of cases smear-positive for TB were 17.0%, 2.5%, and 735.3 per 100,000, respectively. CONCLUSION: The prevalence of TB infection and smear-positive cases of TB increased significantly between 1986 and 2002. Such escalation took place despite the implementation of the national TB programme on this island, which was preceded by a set of special anti-TB measures that resulted in sputum conversion in a substantially large proportion of the smear-positive cases prevalent in the community. The most likely reason for the increase seems to be the absence of a district TB programme with enough efficiency to sustain the gains made from the one-time initial phase of special anti-TB measures. High risk of transmission of TB infection currently observed on this island calls for a drastic and sustained improvement in TB control measures. PMID:15640919

  1. A Novel Reporter Phage To Detect Tuberculosis and Rifampin Resistance in a High-HIV-Burden Population.

    PubMed

    O'Donnell, Max R; Pym, Alexander; Jain, Paras; Munsamy, Vanisha; Wolf, Allison; Karim, Farina; Jacobs, William R; Larsen, Michelle H

    2015-07-01

    Improved diagnostics and drug susceptibility testing for Mycobacterium tuberculosis are urgently needed. We developed a more powerful mycobacteriophage (Φ(2)GFP10) with a fluorescent reporter. Fluorescence-activated cell sorting (FACS) allows for rapid enumeration of metabolically active bacilli after phage infection. We compared the reporter phage assay to GeneXpert MTB/RIF for detection of M. tuberculosis and rifampin (RIF) resistance in sputum. Patients suspected to have tuberculosis were prospectively enrolled in Durban, South Africa. Sputum was incubated with Φ(2)GFP10, in the presence and absence of RIF, and bacilli were enumerated using FACS. Sensitivity and specificity were compared to those of GeneXpert MTB/RIF with an M. tuberculosis culture as the reference standard. A total of 158 patients were prospectively enrolled. Overall sensitivity for M. tuberculosis was 95.90% (95% confidence interval (CI), 90.69% to 98.64%), and specificity was 83.33% (95% CI, 67.18% to 93.59%). In acid-fast bacillus (AFB)-negative sputum, sensitivity was 88.89% (95% CI, 73.92% to 96.82%), and specificity was 83.33% (95% CI, 67.18% to 93.59%). Sensitivity for RIF-resistant M. tuberculosis in AFB-negative sputum was 90.00% (95% CI, 55.46% to 98.34%), and specificity was 91.94% (95% CI, 82.16% to 97.30%). Compared to GeneXpert, the reporter phage was more sensitive in AFB smear-negative sputum, but specificity was lower. The Φ(2)GFP10 reporter phage showed high sensitivity for detection of M. tuberculosis and RIF resistance, including in AFB-negative sputum, and has the potential to improve phenotypic testing for complex drug resistance, paucibacillary sputum, response to treatment, and detection of mixed infection in clinical specimens.

  2. Risk factors for nosocomial tuberculosis transmission among health care workers.

    PubMed

    Ito, Yutaka; Nagao, Miki; Iinuma, Yoshitsugu; Matsumura, Yasufumi; Yamamoto, Masaki; Takakura, Shunji; Igawa, Junko; Yamanaka, Hiroe; Hashimoto, Akiko; Hirai, Toyohiro; Niimi, Akio; Ichiyama, Satoshi; Mishima, Michiaki

    2016-05-01

    We conducted hospital-based contact investigations of 55 serial sputum smear-positive tuberculosis (TB) patients and 771 health care workers (HCWs) from 2006-2013. HCWs who made contact with TB patients in the absence of appropriate airborne precautions were evaluated using interferon gamma release assays to identify TB infection. Twenty-nine HCWs (3.8%) were newly diagnosed with TB infection. The 10 TB patients responsible for transmission had a duration of contact of >7 days by multivariate analysis. PMID:26777287

  3. Pulmonary langerhans cell histiocytosis masquerading as tuberculosis in an infant.

    PubMed

    Senanayake, M P; Mettananda, S; De Silva, M V C

    2011-01-01

    A 4-month-old infant presented with continued fever, unresolving bronchopneumonia and household contact with sputum-smear-positive tuberculosis (TB) and showed marginal improvement on anti-TB chemotherapy. Recurrent pneumothorax prompted the clinical diagnosis of TB to be revised. High-resolution CT scan of the chest and open lung biopsy confirmed the diagnosis of pulmonary Langerhans cell histiocytosis. Treatment with prednisolone and vinblastin resulted in settling of fever and resolution of respiratory symptoms and signs. In communities where the prevalence of TB is high, unusual presentations should prompt consideration of alternative diagnoses. PMID:22041471

  4. Tuberculosis in African Americans: clinical characteristics and outcome.

    PubMed Central

    Funnyé, A. S.; Ganesan, K.; Yoshikawa, T. T.

    1998-01-01

    This study examined the clinical characteristics and outcome of pulmonary tuberculosis in African Americans hospitalized in a teaching hospital in south-central Los Angeles from May 1992 through April 1994. The charts of 41 African Americans with culture-positive Mycobacterium tuberculosis were reviewed. Predisposing factors for pulmonary tuberculosis were identified in nearly half of cases. Cough and fever were the most common symptoms. Seventy-six percent had positive acid-fast bacilli (AFB) smears. Nine patients were human immunodeficiency virus (HIV)-positive, and 6 of 9 HIV-positive patients had positive AFB smears whereas 17 of 19 HIV-negative patients had positive AFB smears. Radiographic changes were not significantly different between HIV-positive and HIV-negative patients. Drug resistance was identified in nine of 31 patients (29%). Eight of 41 patients (19.5%) died, with 2 being drug resistant. Human immunodeficiency virus infection was a major predisposing factor for tuberculosis, and no statistical differences were found in radiographic features or AFB smear positivity between HIV-positive and HIV-negative patients. Drug resistance and mortality were disproportionately high. These results indicate that HIV infection and drug resistance are major problems that predispose for tuberculosis infection and make its treatment difficult. PMID:9510620

  5. Short-term and long-term outcomes following DOTS-based treatment for tuberculosis patients in Burkina Faso

    PubMed Central

    Kouanda, S.; Nikiéma, L.; Ouédraogo, G.; Tiendrébeogo, S.; Sondo, B.; Greenwell, F.; Salomon, J. A.

    2012-01-01

    A cross-sectional study was initiated in Burkina Faso’s National Tuberculosis Programme to confirm successful treatment results within 3 months of completing treatment and to characterise longer-term outcomes 12–24 months after completion. The sample (n = 278) included 91 patients who had completed treatment 0–3 months earlier (‘short-term’ sample) and 187 patients who had completed treatment 12–24 months earlier (‘long-term’ sample). All sputum specimens from the short-term sample were confirmed as negative. In the long-term sample, among 154 patients with available information, 13 (8%) had died, 24 were not traced, and 117 (76%) were interviewed and had sputum examinations, of which 2 (2%) were smear-positive. Recording of successful treatment outcomes shows good validity. PMID:26392942

  6. Seasonality of reported tuberculosis cases from 2006 to 2010 in Wuhan, China.

    PubMed

    Luo, T; Sumi, A; Zhou, D; Kobayashi, N; Mise, K; Yu, B; Kong, D; Wang, J; Duan, Q

    2014-10-01

    We investigated the seasonality of tuberculosis (TB) in Wuhan, China, to evaluate the increased risk of disease transmission during each season and to develop an effective TB control strategy. We applied spectral analysis to the weekly prevalence data of sputum smear positive (SSP) and sputum smear negative (SSN) pulmonary TB reported from 2006 to 2010. Cases of both SSP and SSN feature 1·0- and 0·5-year periodic modes. The least squares method was used to fit curves to the two periodic modes for SSP and SSN data. The curves demonstrated dominant peaks in spring similar to cases reported previously for other locations. Notably for SSP, dominant peaks were also observed in summer. The spring peaks of SSP and SSN were explained in terms of poorly ventilated and humid rooms and vitamin D deficiency. For the summer peaks of SSP, summer influenza epidemics in Wuhan may contribute to the increase in TB prevalence.

  7. Rapid Whole-Genome Sequencing of Mycobacterium tuberculosis Isolates Directly from Clinical Samples

    PubMed Central

    Brown, Amanda C.; Einer-Jensen, Katja; Holdstock, Jolyon; Houniet, Darren T.; Chan, Jacqueline Z. M.; Depledge, Daniel P.; Nikolayevskyy, Vladyslav; Broda, Agnieszka; Stone, Madeline J.; Christiansen, Mette T.; Williams, Rachel; McAndrew, Michael B.; Tutill, Helena; Brown, Julianne; Melzer, Mark; Rosmarin, Caryn; McHugh, Timothy D.; Shorten, Robert J.; Drobniewski, Francis; Speight, Graham; Breuer, Judith

    2015-01-01

    The rapid identification of antimicrobial resistance is essential for effective treatment of highly resistant Mycobacterium tuberculosis. Whole-genome sequencing provides comprehensive data on resistance mutations and strain typing for monitoring transmission, but unlike for conventional molecular tests, this has previously been achievable only from cultures of M. tuberculosis. Here we describe a method utilizing biotinylated RNA baits designed specifically for M. tuberculosis DNA to capture full M. tuberculosis genomes directly from infected sputum samples, allowing whole-genome sequencing without the requirement of culture. This was carried out on 24 smear-positive sputum samples, collected from the United Kingdom and Lithuania where a matched culture sample was available, and 2 samples that had failed to grow in culture. M. tuberculosis sequencing data were obtained directly from all 24 smear-positive culture-positive sputa, of which 20 were of high quality (>20× depth and >90% of the genome covered). Results were compared with those of conventional molecular and culture-based methods, and high levels of concordance between phenotypical resistance and predicted resistance based on genotype were observed. High-quality sequence data were obtained from one smear-positive culture-negative case. This study demonstrated for the first time the successful and accurate sequencing of M. tuberculosis genomes directly from uncultured sputa. Identification of known resistance mutations within a week of sample receipt offers the prospect for personalized rather than empirical treatment of drug-resistant tuberculosis, including the use of antimicrobial-sparing regimens, leading to improved outcomes. PMID:25972414

  8. Household Contact Screening and Yield of Tuberculosis Cases—A Clinic Based Study in Chennai, South India

    PubMed Central

    Nair, Dina; Rajshekhar, Nandita; Klinton, Joel Shyam; Watson, Basilea; Velayutham, Banurekha; Tripathy, Jaya Prasad; Jawahar, Mohideen Shaheed; Swaminathan, Soumya

    2016-01-01

    Background Contact investigation is an active case finding strategy to increase detection of Tuberculosis (TB) and a key component of TB control programs. The household contacts are at a higher risk of exposure than members of the general population. The information on the value and yield of household contact screening and the approaches used in high incidence settings like India is limited. Objective To evaluate the yield of active case finding in household contacts of newly diagnosed smear positive TB patients and the factors associated with increased yield. Method Retrospective record review of the household contacts of newly diagnosed sputum smear positive patients (index case) enrolled in a clinical trial at National Institute of Research in Tuberculosis, Chennai during the period 2007–2014. A sequential screening algorithm with chest x-ray followed by symptom screen was employed to identify presumptive TB patients. Results 643 household contacts of 280 index TB patients were identified out of which 544 (85%) consented for screening. 71/544 (13%) patients had an abnormal chest radiograph and out of them 70% were symptomatic. A total of 29/544 (5.3%) contacts were found to have TB among whom 23/29 (79%) were sputum smear positive. The number needed to screen (NNS) to identify a new TB case among all household contacts was 19 and among those with an abnormal CXR was 02. Age group > 44 years, male gender and siblings of the index case was associated with abnormal chest radiograph whereas age group between 15–44 was significantly associated with developing TB disease among household contacts. Conclusion Active screening among household contacts is an effective way to improve TB case detection. The yield for new TB cases among contacts with abnormal x-ray was high in this study and the use of Chest X-rays in combination with symptom screen is recommended. PMID:27583974

  9. A tuberculosis nationwide prevalence survey in Gambia, 2012

    PubMed Central

    Kendall, Lindsay; Bashorun, Adedapo; Linda, Christopher; Omoleke, Semeeh; Jeffries, David; Maane, Rahmatulai; Alorse, Beatrice Dei; Alorse, William Dei; Okoi, Catherine Bi; Mlaga, Kodjovi D; Kinteh, Ma Ansu; Donkor, Simon; de Jong, Bouke C; Antonio, Martin; d’Alessandro, Umberto

    2016-01-01

    Abstract Objective To estimate the population prevalence of active pulmonary tuberculosis in Gambia. Methods Between December 2011 and January 2013, people aged ≥ 15 years participating in a nationwide, multistage cluster survey were screened for active pulmonary tuberculosis with chest radiography and for tuberculosis symptoms. For diagnostic confirmation, sputum samples were collected from those whose screening were positive and subjected to fluorescence microscopy and liquid tuberculosis cultures. Multiple imputation and inverse probability weighting were used to estimate tuberculosis prevalence. Findings Of 100 678 people enumerated, 55 832 were eligible to participate and 43 100 (77.2%) of those participated. A majority of participants (42 942; 99.6%) were successfully screened for symptoms and by chest X-ray. Only 5948 (13.8%) were eligible for sputum examination, yielding 43 bacteriologically confirmed, 28 definite smear-positive and six probable smear-positive tuberculosis cases. Chest X-ray identified more tuberculosis cases (58/69) than did symptoms alone (43/71). The estimated prevalence of smear-positive and bacteriologically confirmed pulmonary tuberculosis were 90 (95% confidence interval, CI: 53–127) and 212 (95% CI: 152–272) per 100 000 population, respectively. Tuberculosis prevalence was higher in males (333; 95% CI: 233–433) and in the 35–54 year age group (355; 95% CI: 219–490). Conclusion The burden of tuberculosis remains high in Gambia but lower than earlier estimates of 490 per 100 000 population in 2010. Less than half of all cases would have been identified based on smear microscopy results alone. Successful control efforts will require interventions targeting men, increased access to radiography and more accurate, rapid diagnostic tests. PMID:27274595

  10. Burden of tuberculosis in Kampala, Uganda.

    PubMed Central

    Guwatudde, David; Zalwango, Sarah; Kamya, Moses R.; Debanne, Sara M.; Diaz, Mireya I.; Okwera, Alphonse; Mugerwa, Roy D.; King, Charles; Whalen, Christopher C.

    2003-01-01

    OBJECTIVE: To determine the prevalence and incidence of tuberculosis in one of Uganda's poor peri-urban areas. METHODS: Multi-stage sampling was used to select a sample of households whose members were evaluated for presence of signs and/or symptoms of active tuberculosis; history of tuberculosis treatment; and relevant demographic, socioeconomic, and household environment characteristics. Patients with suspected tuberculosis underwent standardized evaluation for active disease. FINDINGS: A sample of 263 households with 1142 individuals was evaluated. Nineteen people were classified as having had tuberculosis during the one-year reference period (May 2001-April 2002): nine (47%) cases already had been diagnosed through the health care system, while 10 cases (53%) were diagnosed through the survey. The prevalences for all forms of tuberculosis and for sputum smear-positive tuberculosis were 14.0 (95% confidence interval (CI) 7.8-20.3) and 4.4 (CI = 0.83-7.89) per thousand, respectively. The incidences for all forms of tuberculosis and for sputum smear-positive tuberculosis were 9.2 (CI = 3.97-14.4) and 3.7 (CI = 0.39-6.95) per thousand per year, respectively. CONCLUSION: The rate of tuberculosis in this peri-urban community was exceptionally high and may be underestimated by current surveillance systems. The need for interventions aimed at reducing tuberculosis transmission in this, and other similar communities with high case rates, is urgent. PMID:14758406

  11. Diagnostic Accuracy of the Small Membrane Filtration Method for Diagnosis of Pulmonary Tuberculosis in a High-HIV-Prevalence Setting.

    PubMed

    Boum, Yap; Kim, Soyeon; Orikiriza, Patrick; Acuña-Villaorduña, Carlos; Vinhas, Solange; Bonnet, Maryline; Nyehangane, Dan; Mwanga-Amumpaire, Juliet; Fennelly, Kevin P; Jones-López, Edward C

    2016-06-01

    Sputum acid-fast bacilli (AFB) smear microscopy has suboptimal sensitivity but remains the most commonly used laboratory test to diagnose pulmonary tuberculosis (TB). We prospectively evaluated the small membrane filtration (SMF) method that concentrates AFB in a smaller area to facilitate detection to improve the diagnostic performance of microscopy. We enrolled adults with suspicion of pulmonary TB from health facilities in southwestern Uganda. Clinical history, physical examination, and 3 sputum samples were obtained for direct fluorescent AFB smear, SMF, Xpert MTB/RIF, and MGIT culture media. Sensitivity and specificity were estimated for SMF, AFB smear, and Xpert MTB/RIF, using MGIT as the reference standard. The analysis was stratified according to HIV status. From September 2012 to April 2014, 737 participants were included in the HIV-infected stratum (146 [20.5%] were culture positive) and 313 were in the HIV-uninfected stratum (85 [28%] were culture positive). In HIV-infected patients, the sensitivity of a single SMF was 67.4% (95% confidence interval [CI], 59.9% to 74.1%); for AFB, 68.0% (95% CI, 60.6% to 74.6%); and for Xpert MTB/RIF, 91.0% (95% CI, 85.0% to 94.8%). In HIV-uninfected patients, the corresponding sensitivities were 72.5% (95% CI, 62.1% to 80.9%), 80.3% (95% CI, 70.8% to 87.2%), and 93.5% (95% CI, 85.7% to 97.2%). The specificity for all 3 tests in both HIV groups was ≥96%. In this setting, the SMF method did not improve the diagnostic accuracy of sputum AFB. The Xpert MTB/RIF assay performed well in both HIV-infected and -uninfected groups.

  12. Diagnostic Accuracy of the Small Membrane Filtration Method for Diagnosis of Pulmonary Tuberculosis in a High-HIV-Prevalence Setting.

    PubMed

    Boum, Yap; Kim, Soyeon; Orikiriza, Patrick; Acuña-Villaorduña, Carlos; Vinhas, Solange; Bonnet, Maryline; Nyehangane, Dan; Mwanga-Amumpaire, Juliet; Fennelly, Kevin P; Jones-López, Edward C

    2016-06-01

    Sputum acid-fast bacilli (AFB) smear microscopy has suboptimal sensitivity but remains the most commonly used laboratory test to diagnose pulmonary tuberculosis (TB). We prospectively evaluated the small membrane filtration (SMF) method that concentrates AFB in a smaller area to facilitate detection to improve the diagnostic performance of microscopy. We enrolled adults with suspicion of pulmonary TB from health facilities in southwestern Uganda. Clinical history, physical examination, and 3 sputum samples were obtained for direct fluorescent AFB smear, SMF, Xpert MTB/RIF, and MGIT culture media. Sensitivity and specificity were estimated for SMF, AFB smear, and Xpert MTB/RIF, using MGIT as the reference standard. The analysis was stratified according to HIV status. From September 2012 to April 2014, 737 participants were included in the HIV-infected stratum (146 [20.5%] were culture positive) and 313 were in the HIV-uninfected stratum (85 [28%] were culture positive). In HIV-infected patients, the sensitivity of a single SMF was 67.4% (95% confidence interval [CI], 59.9% to 74.1%); for AFB, 68.0% (95% CI, 60.6% to 74.6%); and for Xpert MTB/RIF, 91.0% (95% CI, 85.0% to 94.8%). In HIV-uninfected patients, the corresponding sensitivities were 72.5% (95% CI, 62.1% to 80.9%), 80.3% (95% CI, 70.8% to 87.2%), and 93.5% (95% CI, 85.7% to 97.2%). The specificity for all 3 tests in both HIV groups was ≥96%. In this setting, the SMF method did not improve the diagnostic accuracy of sputum AFB. The Xpert MTB/RIF assay performed well in both HIV-infected and -uninfected groups. PMID:27030493

  13. Image processing techniques for identifying Mycobacterium tuberculosis in Ziehl-Neelsen stains.

    PubMed

    Sadaphal, P; Rao, J; Comstock, G W; Beg, M F

    2008-05-01

    Worldwide, laboratory technicians tediously read sputum smears for tuberculosis (TB) diagnosis. We demonstrate proof of principle of an innovative computational algorithm that successfully recognizes Ziehl-Neelsen (ZN) stained acid-fast bacilli (AFB) in digital images. Automated, multi-stage, color-based Bayesian segmentation identified possible 'TB objects', removed artifacts by shape comparison and color-labeled objects as 'definite', 'possible' or 'non-TB', bypassing photomicrographic calibration. Superimposed AFB clusters, extreme stain variation and low depth of field were challenges. Our novel method facilitates electronic diagnosis of TB, permitting wider application in developing countries where fluorescent microscopy is currently inaccessible and unaffordable. We plan refinement and validation in the future.

  14. Population Based National Tuberculosis Prevalence Survey among Adults (>15 Years) in Pakistan, 2010–2011

    PubMed Central

    Qadeer, Ejaz; Fatima, Razia; Yaqoob, Aashifa; Tahseen, Sabira; Ul Haq, Mahboob; Ghafoor, Abdul; Asif, Muhammad; Straetemans, Masja; Tiemersma, Edine W.

    2016-01-01

    Background We aimed to determine the prevalence of pulmonary tuberculosis (TB) amongst the adult population in 2010–2011 in Pakistan. Method A nationwide cross-sectional survey with multistage cluster sampling was conducted among adults (≥15 years) in 95 clusters in 2010–2011. All consenting participants were screened for cough and by chest X-ray. Participants with presumptive TB submitted two sputum samples for smear microscopy, culture, and molecular testing if needed. The TB prevalence estimates were adjusted for missing data and the cluster design. Result Of 131,329 eligible individuals, 105,913 (81%) participated in the survey, of whom 10,471 (9.9%) were eligible for sputum examination. We found 341 bacteriologically positive TB cases of whom 233 had sputum smear-positive TB. The adjusted prevalence estimates for smear and bacteriologically positive TB were 270/100,000 (95% confidence interval (CI) 217–323), and 398/100,000 (95% CI 333–463), respectively. Only 61% of the diagnosed TB cases screened positive on symptoms (cough >2wks), whereas the other TB cases were detected based on X-ray abnormalities. The TB prevalence increased with age and was 1.8 times higher among men than women. The prevalence-to-notification ratio of smear-positive TB was 3.1 (95% CI 2.5–3.7), was higher among men than women, and increased with age. Conclusion Our data suggest that there is under-detection and/or -notification of TB, especially among men and elderly. TB control should be strengthened specifically in these risk groups. X-ray examination should be combined with symptom screening to enhance case detection. PMID:26863617

  15. Automatic sputum color image segmentation for tuberculosis diagnosis

    NASA Astrophysics Data System (ADS)

    Forero-Vargas, Manuel G.; Sierra-Ballen, Eduard L.; Alvarez-Borrego, Josue; Pech-Pacheco, Jose L.; Cristobal-Perez, Gabriel; Alcala, Luis; Desco, Manuel

    2001-11-01

    Tuberculosis (TB) and other mycobacteriosis are serious illnesses which control is mainly based on presumptive diagnosis. Besides of clinical suspicion, the diagnosis of mycobacteriosis must be done through genus specific smears of clinical specimens. However, these techniques lack of sensitivity and consequently clinicians must wait culture results as much as two months. Computer analysis of digital images from these smears could improve sensitivity of the test and, moreover, decrease workload of the micobacteriologist. Bacteria segmentation of particular species entails a complex process. Bacteria shape is not enough as a discriminant feature, because there are many species that share the same shape. Therefore the segmentation procedure requires to be improved using the color image information. In this paper we present two segmentation procedures based on fuzzy rules and phase-only correlation techniques respectively that will provide the basis of a future automatic particle' screening.

  16. ASCORBID ACID IS DECREASED IN INDUCED SPUTUM OF MILD ASTHMATICS

    EPA Science Inventory

    ABSTRACT
    Evidence suggests that the antioxidant ascorbic acid (AA), plays an essential role in defending against oxidant attack in the airways. Decreased levels of AA have been reported in asthmatics but not at the site directly proximal to asthma pathology, i.e. the bronchial...

  17. ASCORBIC ACID IS DECREASED IN INDUCED SPUTUM OF MILD ASTHMATICS

    EPA Science Inventory

    Asthma is primarily an airways inflammatory disease, and the bronchial airways have been shown to be particularly susceptible to oxidant-induced tissue damage. The antioxidant ascorbic acid (AA) plays an essential role in defending against oxidant attack in the airways. Decreased...

  18. Value of Urine Lipoarabinomannan Grade and Second Test for Optimizing Clinic-based Screening for HIV-associated Pulmonary Tuberculosis

    PubMed Central

    Drain, Paul K.; Losina, Elena; Coleman, Sharon M.; Giddy, Janet; Ross, Douglas; Katz, Jeffrey N.; Bassett, Ingrid V.

    2014-01-01

    Background We assessed the role of urine LAM (lipoarabinomannan) grade and a second LAM test for HIV-associated pulmonary tuberculosis (TB) screening in outpatient clinics in South Africa. Methods We enrolled newly-diagnosed HIV-infected adults (≥18 years) at 4 clinics, excluding those on TB therapy. Participants provided sputum for AFB microscopy and culture. Nurses conducted two rapid urine LAM tests at the point-of-care, and graded positive results from low (“faint”) to high (5+). Culture-confirmed pulmonary TB was the gold standard. We used area under receiver operating curves (AUROC) to compare screening strategies. Results Among 320 HIV-infected adults, median CD4 was 248/mm3 (IQR 107–379/mm3); 54 (17%) were TB culture-positive. 52 (16%) of all participants were LAM-positive by either test; correlation between LAM tests was high. Among 10 “faint” positive results, 2 (20%) had culture-positive TB. Using ≥1+ LAM grade as positive, one LAM test had sensitivity of 41% (95% CI 28–55%) and specificity of 92% (95% CI 88–95%). A 2 LAM test strategy had a sensitivity of 43% (95% CI 29–57%). One LAM test ≥1+ grade (AUROC=0.66; 95% CI 0.60–0.73) was significantly better than sputum AFB alone. The optimal strategy was sequentially performing one LAM test followed by sputum AFB if LAM grade <1+ (AUROC=0.70; 95% CI 0.63–0.77), which had sensitivity of 48% (95% CI 34–62%) and specificity of 91% (95% CI 87–94%). Conclusions In this clinic-based study, “faint” line was a false-positive, second urine LAM test added no value, and an optimal screening strategy was one LAM test followed by sputum AFB microscopy for urine LAM-negative people. PMID:25415288

  19. Improved recovery of mycobacteria from respiratory secretions of patients with cystic fibrosis.

    PubMed

    Whittier, S; Hopfer, R L; Knowles, M R; Gilligan, P H

    1993-04-01

    Pulmonary colonization and infection of patients with cystic fibrosis by Mycobacterium spp. has recently been recognized as a potentially important clinical problem. However, frequent contamination of mycobacterial cultures by pseudomonads has hampered efforts to define the extent of this problem. This study was done to evaluate current techniques and to establish a more efficient method of recovering mycobacteria from respiratory secretions of patients with cystic fibrosis. Decontamination of respiratory specimens (n = 121) with 0.25% N-acetyl-L-cysteine and 1% sodium hydroxide (NALC-NaOH) was associated with a high rate of pseudomonas overgrowth for both Lowenstein-Jensen slants (74%) and BacTec vials supplemented with PANTA (polymyxin B [50 U/ml], amphotericin B [5 micrograms/ml], nalidixic acid [20 micrograms/ml], trimethoprim [5 micrograms/ml], azlocillin [10 micrograms/ml]) (36%). This overgrowth limited recovery of mycobacteria to only 64% (9 of 14) of specimens positive by smear for acid-fast bacilli (AFB). Decontamination of specimens (n = 441) with NALC-NaOH, followed by 5% oxalic acid treatment, resulted in contamination of only 5% of Lowenstein-Jensen slants and 3% of BacTec vials. AFB were recovered from all 90 AFB smear-positive specimens following the use of this decontamination technique. We recommend that respiratory secretions be decontaminated with NALC-NaOH and oxalic acid to decrease the incidence of Pseudomonas aeruginosa overgrowth. PMID:8463398

  20. Intensive Tuberculosis Screening for HIV-Infected Patients Starting ART in Durban, South Africa

    PubMed Central

    Bassett, Ingrid V.; Wang, Bingxia; Chetty, Senica; Giddy, Janet; Losina, Elena; Mazibuko, Matilda; Bearnot, Benjamin; Allen, Jenny; Tech, B; Walensky, Rochelle P.; Freedberg, Kenneth A.

    2011-01-01

    Background The World Health Organization (WHO) recommends cough as the trigger for tuberculosis (TB) screening in HIV-infected patients, with acid fast bacillus (AFB) smear as the initial diagnostic test. Our objective was to assess the yield and cost of a more intensive TB screening in HIV-infected patients starting antiretroviral therapy (ART) in Durban, South Africa. Methods We prospectively enrolled adults, regardless of TB signs/symptoms, undergoing pre-ART training from May ‘07–May ‘08. Following symptom screen, patients expectorated sputum for AFB smear, TB polymerase chain reaction (PCR), and mycobacterial culture. Sensitivity and specificity of different symptoms and tests, alone and in combination, were compared to a gold standard of 6-week TB culture results. Program costs included personnel, materials and cultures. Results Of 1,035 subjects, 487 (59%) were female; median CD4 count was 100/μl. Two-hundred and ten (20%) were receiving TB treatment and were excluded. Of the remaining 825 subjects, 158 (19%) had positive sputum cultures, of whom 14 (9%) had a positive AFB smear and 82 (52%) reported cough. The combination of cough, other symptoms, AFB smear, and chest x-ray had 93% (CI 88–97%) sensitivity and 15% (CI 13–18%) specificity. The incremental cost of intensive screening including culture was $360/additional TB case identified. Conclusions Nearly 20% of patients starting ART in Durban, South Africa had undiagnosed, culture-positive pulmonary TB. Despite WHO recommendations, neither cough nor AFB smear were adequately sensitive for screening. TB sputum cultures should be performed before ART initiation, regardless of symptoms, in areas of high HIV/TB prevalence. PMID:20735240

  1. Treatability study for Hill AFB`s Operable Unit-1: Enhanced microaerobic dechlorination using various electron donors. MasMajor report

    SciTech Connect

    Breed, P.G.

    1999-05-13

    A treatability study of the microaerobic biodegradation of cis-dichloroethene (c-DCE) was completed using a series of eight continuously operated columns filled with contaminated soils from Hill Air Force Base`s Operable Unit 1. Columns were supplied groundwater from the site, vitamins and yeast, and an electron donor solution containing one of the following donors: n-butyric acid, benzoic acid, lactic acid, propionic acid, n-propanol, or toluene. Concentrations of c-DCE varied over six months and ranged from 2736 micrograms/L to 30 micrograms/L. Though attempted as an anaerobic study, the ability to continuously eliminate oxygen from an active system proved difficult and columns operated as microaerobic systems. In all columns the degradation of c-DCE was observed, however, the removal efficiencies determined by comparing the influent and effluent concentrations were highly inconsistent throughout the experiment. By comparing the background columns to the columns supplied electron donors, it does not appear the addition of vitamins or electron donors enhance the indigenous microorganism`s ability to remove c-DCE. While c-DCE removal within the background column averaged 17%, the vitamin amended control column averaged only 7% c-DCE removal within the column and the electron donor supplied columns averaged between 7% removal and 5% apparent production. Of the electron donors supporting c-DCE removal, benzoic acid demonstrated 7% removal followed closely by propionic acid and n-propanol, both showing 5% c-DCE removal.

  2. Comparative study of clinico-bacterio-radiological profile and treatment outcome of smokers and nonsmokers suffering from pulmonary tuberculosis

    PubMed Central

    Rathee, Deepti; Arora, Piyush; Meena, Manoj; Sarin, Rohit; Chakraborty, Pitambar; Jaiswal, Anand; Goyal, Mukesh

    2016-01-01

    Introduction: Tuberculosis (TB) is one of the leading causes of death and disease worldwide. Tobacco smoking has been linked as a risk factor for TB. This study was aimed to affirm the strength of association between smoking and pulmonary TB. Materials and Methods: Pulmonary TB patients aged between 18 and 65 years were enrolled and followed-up until treatment completion. Two consecutive sputum smears were examined from each patient for the presence of acid-fast bacilli (AFB) using Ziehl–Neelsen technique. Radiological severity of disease was assessed using guidelines of National TB Association of USA. Sputum smears for AFB were graded for positivity as per WHO Revised National TB Control Programme criteria. Response was determined in terms of sputum conversion at the end of intensive phase and final treatment outcomes. Results: Sputum smear grading of 3+ increased from 12.5% to 68.18% and 66.66% as smoking index increased from <100 to 100–299 and >300 (P < 0.05). In nonsmokers, 79.2% patients had minimal disease while only 4.2% had advanced disease as compared to smokers where 52.4% had moderate disease, 26.2% advanced disease, and 21.4% minimal disease (P < 0.01). Smokers had significantly lower treatment success rate (69%) as against nonsmokers and former smokers (93.8% and 90.9%, respectively, P = 0.001) owing to a higher default rate among smokers (28.5%) than nonsmokers (6.3%) and former smokers (9.1%). Conclusion: Smokers during initial presentation, as well as at end of the treatment demonstrate more radiological findings, cavitary disease, and worse sputum AFB smear grading. Smokers also have a poorer treatment success rate largely due to high percentage of default rate thus suggesting noncompliance as a main confounder to treatment success. Focus needs to be made to reduce defaulters which are more common among smokers.

  3. Comparative study of clinico-bacterio-radiological profile and treatment outcome of smokers and nonsmokers suffering from pulmonary tuberculosis

    PubMed Central

    Rathee, Deepti; Arora, Piyush; Meena, Manoj; Sarin, Rohit; Chakraborty, Pitambar; Jaiswal, Anand; Goyal, Mukesh

    2016-01-01

    Introduction: Tuberculosis (TB) is one of the leading causes of death and disease worldwide. Tobacco smoking has been linked as a risk factor for TB. This study was aimed to affirm the strength of association between smoking and pulmonary TB. Materials and Methods: Pulmonary TB patients aged between 18 and 65 years were enrolled and followed-up until treatment completion. Two consecutive sputum smears were examined from each patient for the presence of acid-fast bacilli (AFB) using Ziehl–Neelsen technique. Radiological severity of disease was assessed using guidelines of National TB Association of USA. Sputum smears for AFB were graded for positivity as per WHO Revised National TB Control Programme criteria. Response was determined in terms of sputum conversion at the end of intensive phase and final treatment outcomes. Results: Sputum smear grading of 3+ increased from 12.5% to 68.18% and 66.66% as smoking index increased from <100 to 100–299 and >300 (P < 0.05). In nonsmokers, 79.2% patients had minimal disease while only 4.2% had advanced disease as compared to smokers where 52.4% had moderate disease, 26.2% advanced disease, and 21.4% minimal disease (P < 0.01). Smokers had significantly lower treatment success rate (69%) as against nonsmokers and former smokers (93.8% and 90.9%, respectively, P = 0.001) owing to a higher default rate among smokers (28.5%) than nonsmokers (6.3%) and former smokers (9.1%). Conclusion: Smokers during initial presentation, as well as at end of the treatment demonstrate more radiological findings, cavitary disease, and worse sputum AFB smear grading. Smokers also have a poorer treatment success rate largely due to high percentage of default rate thus suggesting noncompliance as a main confounder to treatment success. Focus needs to be made to reduce defaulters which are more common among smokers. PMID:27625444

  4. Factors Associated with Unfavorable Treatment Outcomes in New and Previously Treated TB Patients in Uzbekistan: A Five Year Countrywide Study

    PubMed Central

    Gadoev, Jamshid; Asadov, Damin; Tillashaykhov, Mirzagolib; Tayler-Smith, Katie; Isaakidis, Petros; Dadu, Andrei; de Colombani, Pierpaolo; Gudmund Hinderaker, Sven; Parpieva, Nargiza; Ulmasova, Dilrabo; Jalolov, Avazbek; Hamraev, Atadjan; Ali, Engy; van den Boom, Martin; Hammerich, Asmus; Gozalov, Ogtay; Dara, Masoud

    2015-01-01

    Background TB is one of the main health priorities in Uzbekistan and relatively high rates of unfavorable treatment outcomes have recently been reported. This requires closer analysis to explain the reasons and recommend interventions to improve the situation. Thus, by using countrywide data this study sought to determine trends in unfavorable outcomes (lost-to-follow-ups, deaths and treatment failures) and describe their associations with socio-demographic and clinical factors. Method A countrywide retrospective cohort study of all new and previously treated TB patients registered in the National Tuberculosis programme between January 2006 and December 2010. Results Among 107,380 registered patients, 67% were adults, with smaller proportions of children (10%), adolescents (4%) and elderly patients (19%). Sixty per cent were male, 66% lived in rural areas, 1% were HIV-infected and 1% had a history of imprisonment. Pulmonary TB (PTB) was present in 77%, of which 43% were smear-positive and 53% were smear-negative. Overall, 83% of patients were successfully treated, 6% died, 6% were lost-to-follow-up, 3% failed treatment and 2% transferred out. Factors associated with death included being above 55 years of age, HIV-positive, sputum smear positive, previously treated, jobless and living in certain provinces. Factors associated with lost-to-follow-up were being male, previously treated, jobless, living in an urban area, and living in certain provinces. Having smear-positive PTB, being an adolescent, being urban population, being HIV-negative, previously treated, jobless and residing in particular provinces were associated with treatment failure. Conclusion Overall, 83% treatment success rate was achieved. However, our study findings highlight the need to improve TB services for certain vulnerable groups and in specific areas of the country. They also emphasize the need to develop unified monitoring and evaluation tools for drug-susceptible and drug-resistant TB, and

  5. Immunogenicity of mycobacterial vesicles in humans: identification of a new tuberculosis antibody biomarker.

    PubMed

    Ziegenbalg, Anke; Prados-Rosales, Rafael; Jenny-Avital, Elisabeth R; Kim, Ryung S; Casadevall, Arturo; Achkar, Jacqueline M

    2013-07-01

    Biomarkers for active tuberculosis (TB) are urgently needed. Mycobacteria produce membrane vesicles (MVs) that contain concentrated immune-modulatory factors that are released into the host. We evaluated the human immune responses to BCG and Mycobacterium tuberculosis MVs to characterize the antibody responses and identify potentially novel TB biomarkers. Serological responses to MVs were evaluated by ELISAs and immunoblots with sera from 16 sputum smear-positive, 12 smear-negative HIV uninfected pulmonary TB patients and 16 BCG vaccinated Tuberculin skin-test positive controls with and without latent tuberculosis infection. MVs from both BCG and M. tuberculosis induced similar responses and were strongly immunogenic in TB patients but not in controls. Several MV-associated antigens appear to induce robust antibody responses, in particular the arabinomanan portion of the cell wall glycolipid lipoarabinomannan. Three proteins at ≈ 36, 25, and 23 kDa were simultaneously recognized by sera from 16/16 smear-positive, 9/12 smear-negative TB patients and 0/16 controls. These results provide promise and encouragement that antibody responses to proteins enriched in MVs of pathogenic mycobacteria may constitute a novel TB biomarker signature that could have diagnostic information.

  6. Improvement of tuberculosis laboratory capacity on Pemba Island, Zanzibar: a health cooperation project.

    PubMed

    Paglia, Maria G; Bevilacqua, Nazario; Haji, Haji Said; Vairo, Francesco; Girardi, Enrico; Nicastri, Emanuele; Muhsin, Juma; Racalbuto, Vincenzo; Jiddawi, Mohammed S; Ippolito, Giuseppe

    2012-01-01

    Low-income countries with high Tuberculosis burden have few reference laboratories able to perform TB culture. In 2006, the Zanzibar National TB Control Programme planned to decentralize TB diagnostics. The Italian Cooperation Agency with the scientific support of the "L. Spallanzani" National Institute for Infectious Diseases sustained the project through the implementation of a TB reference laboratory in a low-income country with a high prevalence of TB. The implementation steps were: 1) TB laboratory design according to the WHO standards; 2) laboratory equipment and reagent supplies for microscopy, cultures, and identification; 3) on-the-job training of the local staff; 4) web- and telemedicine-based supervision. From April 2007 to December 2010, 921 sputum samples were received from 40 peripheral laboratories: 120 TB cases were diagnosed. Of all the smear-positive cases, 74.2% were culture-positive. During the year 2010, the smear positive to culture positive rate increased up to 100%. In March 20, 2010 the Ministry of Health and Social Welfare of Zanzibar officially recognized the Public Health Laboratory- Ivo de Carneri as the National TB Reference Laboratory for the Zanzibar Archipelago. An advanced TB laboratory can represent a low cost solution to strengthen the TB diagnosis, to provide capacity building and mid-term sustainability.

  7. Effectiveness of a community-based observation of anti-tuberculosis treatment in Bangalore City, India, 2010-2011

    PubMed Central

    Kumar, P.; Sagili, K. D.; Enarson, D. A.

    2013-01-01

    Setting: The Revised National Tuberculosis Control Programme in an urban setting of Bangalore City, India. Objectives: To compare treatment outcomes and smear conversion rates among new smear-positive tuberculosis (TB) patients undergoing treatment administered by community directly observed treatment (DOT) providers with those undergoing treatment administered by institutional DOT providers in Bangalore City in 2010-2011. Method: Cohort study of routine data recorded from treatment cards of TB patients undergoing treatment under the public health services from 1 October 2010 to 30 September 2011. Result: Treatment records of 1864 new smear-positive TB patients registered during this period were evaluated. Among those evaluated, 604 (32%) had been administered treatment by community DOT providers and the remainder by institutional DOT providers. The treatment success rate in those undergoing community DOT was 93% (n = 564) and that of those undergoing institutional DOT was 75% (n = 951; RR 1.23, 95%CI 1.19-1.28). The sputum smear conversion rate of patients who underwent community DOT was 92% and that of those who underwent institutional DOT was 71% at the end of 2 months. Conclusion: We conclude that community DOT for treatment supervision of TB patients is more effective than institutional DOT and that it should be reinforced. PMID:26393035

  8. Engaging private providers and Ayurvedic practitioners in Bilaspur, India: did it increase TB case detection?

    PubMed

    Bhardwaj, R R; Oeltmann, J E; Ravichandra, C; Chadda, V K; Das, M; Kumar, A M V

    2016-06-21

    To find 'missing' tuberculosis (TB) cases, in November 2014 we trained private practitioners (PPs) and Ayurvedic practitioners (APs; Indian system of medicine) in Bilaspur district, India, to identify patients with presumptive TB and refer them to sputum microscopy centres. To reinforce this training, we sent weekly text message reminders during January-March 2015. All 50 APs and 23 of 29 PPs participated. The number of patients with presumptive TB referred by the PPs and APs increased from 38 (January-March 2014) to 104 (January-March 2015), and the number of smear-positive TB patients diagnosed increased from 5 to 16, a 220% increase. While the intervention increased the number of referrals, it did not impact case detection at district level, due to the short duration of the intervention and the non-dominant private sector.

  9. Shortfalls identified in the management of tuberculosis for Mozambican patients obtaining health care services in Malawi.

    PubMed

    Muula, Adamson S; Nyasulu, Yohane; Feluzi, Henry; Magalasi, Collins

    2003-06-01

    We report findings of a pilot qualitative study in which we aimed to determine management gaps among TB patients from Mozambique obtaining health care services in Malawi. The study was conducted between April and May 2002 involved twelve health workers and 4 Mozambican patients. Semi-structured questionnaires were used and responses were followed up with in-depth interviews. Several areas of management gaps were identified. These included; language barrier if patients are formally referred with documents in Portuguese; lack of follow-up system in case of patients defaulting; no structured contact-tracing possibilities and no initiation of Isoniazid prophylaxis in the case of children living in households with a sputum smear positive adult case. We conclude that logistical management gaps exist in the management of TB patients from Mozambique obtaining care in Malawian health care facilities.

  10. Rapid Direct Testing of Susceptibility of Mycobacterium tuberculosis to Isoniazid and Rifampin on Nutrient and Blood Agar in Resource-Starved Settings

    PubMed Central

    Ikram, Aamer; Coban, Ahmet Yilmaz; Martin, Anandi

    2012-01-01

    In this study, we evaluated the performance of blood agar (by macroscopic growth) and nutrient agar (by a microcolony detection method) for drug susceptibility testing of Mycobacterium tuberculosis against rifampin (RIF) and isoniazid (INH), using 67 smear-positive sputum specimens. The direct proportion method on Lowenstein-Jensen (LJ) medium was used as the “gold standard.” Compared with LJ medium, results for both media were in 100% agreement for RIF, while for INH the agreement levels for blood agar and nutrient agar were 98% and 95%, respectively. Within 2 weeks, 100% of specimens yielded results on blood agar, while 96.8% of specimens yielded results on nutrient agar. Our study showed that blood agar and nutrient agar can be used as alternative media for direct susceptibility testing of RIF and INH, especially in resource-poor settings. PMID:22357498

  11. Engaging private providers and Ayurvedic practitioners in Bilaspur, India: did it increase TB case detection?

    PubMed Central

    Oeltmann, J. E.; Ravichandra, C.; Chadda, V. K.; Das, M.; Kumar, A. M. V.

    2016-01-01

    To find ‘missing’ tuberculosis (TB) cases, in November 2014 we trained private practitioners (PPs) and Ayurvedic practitioners (APs; Indian system of medicine) in Bilaspur district, India, to identify patients with presumptive TB and refer them to sputum microscopy centres. To reinforce this training, we sent weekly text message reminders during January–March 2015. All 50 APs and 23 of 29 PPs participated. The number of patients with presumptive TB referred by the PPs and APs increased from 38 (January–March 2014) to 104 (January–March 2015), and the number of smear-positive TB patients diagnosed increased from 5 to 16, a 220% increase. While the intervention increased the number of referrals, it did not impact case detection at district level, due to the short duration of the intervention and the non-dominant private sector. PMID:27358811

  12. Engaging private providers and Ayurvedic practitioners in Bilaspur, India: did it increase TB case detection?

    PubMed

    Bhardwaj, R R; Oeltmann, J E; Ravichandra, C; Chadda, V K; Das, M; Kumar, A M V

    2016-06-21

    To find 'missing' tuberculosis (TB) cases, in November 2014 we trained private practitioners (PPs) and Ayurvedic practitioners (APs; Indian system of medicine) in Bilaspur district, India, to identify patients with presumptive TB and refer them to sputum microscopy centres. To reinforce this training, we sent weekly text message reminders during January-March 2015. All 50 APs and 23 of 29 PPs participated. The number of patients with presumptive TB referred by the PPs and APs increased from 38 (January-March 2014) to 104 (January-March 2015), and the number of smear-positive TB patients diagnosed increased from 5 to 16, a 220% increase. While the intervention increased the number of referrals, it did not impact case detection at district level, due to the short duration of the intervention and the non-dominant private sector. PMID:27358811

  13. Rapid direct testing of susceptibility of Mycobacterium tuberculosis to isoniazid and rifampin on nutrient and blood agar in resource-starved settings.

    PubMed

    Satti, Luqman; Ikram, Aamer; Coban, Ahmet Yilmaz; Martin, Anandi

    2012-05-01

    In this study, we evaluated the performance of blood agar (by macroscopic growth) and nutrient agar (by a microcolony detection method) for drug susceptibility testing of Mycobacterium tuberculosis against rifampin (RIF) and isoniazid (INH), using 67 smear-positive sputum specimens. The direct proportion method on Lowenstein-Jensen (LJ) medium was used as the "gold standard." Compared with LJ medium, results for both media were in 100% agreement for RIF, while for INH the agreement levels for blood agar and nutrient agar were 98% and 95%, respectively. Within 2 weeks, 100% of specimens yielded results on blood agar, while 96.8% of specimens yielded results on nutrient agar. Our study showed that blood agar and nutrient agar can be used as alternative media for direct susceptibility testing of RIF and INH, especially in resource-poor settings.

  14. Tuberculosis case--finding and treatment in the central prison of Qazvin province, Islamic Republic of Iran.

    PubMed

    Assefzadeh, M; Barghi, R Gh; Shahidi, Sh S

    2009-01-01

    As a part of a comprehensive programme of tuberculosis (TB) control in the central prison in Qazvin, Islamic Republic of Iran, a programme of active case-finding was carried out from February 2004 to July 2005. From the 768 prisoners examined, 41 (5.3%) were suspected of TB and gave sputum samples. A total of 7 smear-positive TB cases were found, giving a TB prevalence in the prison of 910 per 100,000, 113 times the total TB prevalence in Qazvin province in the same year. From 7 diagnosed and treated patients, 4 were cured with the category 1 standard regimen, 1 completed treatment and 2 failed to complete treatment. Improvements are needed in TB case recognition of prisoners, especially newcomers.

  15. Soil Flushing Through a Thick Vadose Zone: Perchlorate Removal Documented at Edwards AFB, California

    NASA Astrophysics Data System (ADS)

    Battey, T. F.; Shepard, A. J.; Tait, R. J.

    2007-12-01

    There are currently few viable alternatives for perchlorate remediation in the vadose zone, particularly for the relatively thick vadose zones that are typical in the arid southwest where many perchlorate sites occur. Perchlorate in the vadose zone occurs in the form of highly soluble salts that may represent a risk to human or ecological receptors, and may also represent a threat to the underlying groundwater. A soil flushing treatability study was conducted at Edwards Air Force Base in the Mojave Desert of southern California at a site with a 129-foot thick vadose zone consisting primarily of clayey sand. This study utilized an infiltration gallery in conjunction with extraction, treatment, and re-injection of groundwater at the site, which contained perchlorate-contaminated soil and groundwater. The study objective was to evaluate the effectiveness of the infiltration gallery to 1) introduce treated groundwater back into the aquifer and 2) wash the perchlorate from the vadose zone soils to the aquifer. The infiltration gallery consisted of slotted PVC pipes within a highly permeable engineered bed of washed gravel. The initial water introduced into the gallery was amended with potassium bromide tracer. A downhole neutron probe was used to track the movement of the wetting front downward and outward from the gallery. Successive neutron measurements in vertical access tubes revealed that the introduced water reached the 125-foot bottom of the access tubes 14 weeks after the water was introduced into the gallery. The bromide tracer was detected in groundwater immediately below the gallery approximately 1 week later. The infiltration gallery was able to sustain an average flow rate of 2.3 gallons per minute. Prior to infiltration, the perchlorate concentration in groundwater below the gallery was 4,500 µg/L. Approximately 18 weeks after the start of infiltration, a perchlorate spike of 72,400 µg/L was detected below the gallery. The increase in perchlorate groundwater concentrations indicates the transfer of perchlorate from the vadose zone to the saturated zone, where it was readily captured by an adjacent groundwater extraction well. Continued flushing of treated water through the vadose and saturated zones resulted in a rapid decline in perchlorate groundwater concentrations. Confirmation soil boreholes documented the effectiveness of the soil treatment after perchlorate groundwater concentrations returned to their pre-soil flushing levels. This treatability study demonstrates that perchlorate can be removed from a thick sandy vadose zone by controlled infiltration with associated hydraulic control of groundwater to capture the leached perchlorate. The treatability study results also indicates that an infiltration gallery may provide 1) a cost-effective alternative to injection wells for reintroducing treated groundwater to the aquifer and 2) an effective mechanism for the delivery of amendments to the vadose zone and aquifer for promoting enhanced biodegradation of perchlorate in soil and groundwater.

  16. Remedial investigation/feasibility study analysis asphalt storage area, Elmendorf AFB, Alaska. Master's thesis

    SciTech Connect

    Miller, N.S.

    1993-01-01

    This report is focused on an abandoned material storage area located on Elmendorf Air Force Base (EAFB), Alaska. The site is located approximately 2000 feet from the east end of the east/west runway and includes approximately 25 acres. The site was used for asphalt storage and preparation activities during the 1940s and 1950s. Approximately 4,500 drums of asphalt and 29 drums of unknown materials have been abandoned at the site. The drums are located in 32 areas throughout the 25-acre site. Following several decades of exposure to the elements, many of the drums have corroded and leaked to the ground surface. Several acres of soil are inundated with liquid asphalt that has leaked from the drums. Depths of the asphalt range from 6 to 10 inches in areas where surface anomalies have created depressions, and thus a collection point for the asphalt. A 14-x 18-x 4 foot wood frame pit used to support previous asphalt operations is located at the north end of the site. The pit contains approximately 2300 gallons of asphalt. There are also locations where the soil appears to be contaminated by petroleum products other than asphalt.

  17. 78 FR 48292 - Amendment of Class D Airspace; Grand Forks AFB, ND

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-08

    ... (78 FR 33016) Docket No. FAA-2013-0261. Interested parties were ] invited to participate in this... 12866; (2) is not a ``significant rule'' under DOT Regulatory Policies and Procedures (44 FR 11034... U.S.C. 106(g), 40103, 40113, 40120; E.O. 10854, 24 FR 9565, 3 CFR, 1959-1963 Comp., p. 389. Sec....

  18. 1987 ASCAN Readdy in ejection seat during training at Vance AFB, Oklahoma

    NASA Technical Reports Server (NTRS)

    1987-01-01

    William F. Readdy, 1987 Astronaut Candidate (ASCAN) and member of Astronaut Class 12 (Group 12), straps himself into ejection seat as personnel look on during training at Vance Air Force Base (VAFB), Enid, Oklahoma. The new group of ASCANs participated in parachute training while at VAFB, 09-09-87 through 09-11-87.

  19. ASCAN Susan J. Helms participates in ejection seat training at Vance AFB

    NASA Technical Reports Server (NTRS)

    1990-01-01

    1990 Group 13 Astronaut Candidate (ASCAN) Susan J. Helms participates in ejection seat training at Vance Air Force Base (VAFB), Enid, Oklahoma. Wearing a helmet and strapped into a seat, Helms will experience the 'feel' of emergency ejection from a jet aircraft during this test. Helms, along with the rest of the 1990 Astronaut Class, was part of ground egress and parasail training exercises at VAFB from 07-29-90 through 07-31-90. Photograph was taken by NASA JSC contract photographer Robert L. Walck.

  20. ARAC dispersion modeling support for January--March 1995 Vandenberg AFB launches

    SciTech Connect

    Baskett, R.L.; Pace, J.C.

    1995-05-01

    The Glory Trip (GT) 17-PA Peacekeeper launch originally scheduled at Vandenberg Air Force Base (VAFB) between 15 and 20 November 1994 was cancelled based on modeled toxic exhaust cloud calculations. The Missile Flight Control Branch, 30th Space Wing Safety Office (30 SW/SEY), made several successive ``No Go`` decisions using Version 7. 05 Rocket Exhaust Effluent Dispersion Model (REEDM) with forecasted meteorological conditions. REEDM runs made from T-14 hours to T-30 minutes predicted that ground-level concentrations of hydrogen chloride (HCl) gas from the catastrophic abort case would exceed 5 ppM, the ``instantaneous`` ambient air concentration ``Tier 2`` limit at that time, modeled as a peak 1-minute cloud centerline concentration. Depending on the forecasted wind direction and speed at launch time, this limit was predicted to be exceeded sometimes at Base Housing, approximately 10 km southeast of the launch, and during other launch windows at the town of Casmalia, about 5 km east- southeast. In late December 1994, the LLNL Atmospheric Release Advisory Capability (ARAC) program modeled the aborted November 1994 Peacekeeper launch and compared its results with REEDM. This initial comparison showed that the ARAC model predicted values about 1/3 as large as REEDM for the limiting case at Base Housing. Subsequently ARAC was asked to provide real-time modeling support to 30 SW/SEY during the rescheduled Peacekeeper GT 17-PA launch in January 1995 and two Minuteman launches in February and March. This report first briefly discusses the model differences and then summarizes the results of the three supported launches.

  1. 78 FR 17085 - Amendment of Multiple Restricted Areas; Eglin AFB, FL

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-20

    ...'' under DOT Regulatory Policies and Procedures (44 FR 11034; February 26, 1979); and (3) does not warrant... follows: Authority: 49 U.S.C. 106(g), 40103, 40113, 40120; E.O. 10854, 24 FR 9565, 3 CFR, 1959-1963 Comp... Federal Aviation Administration 14 CFR Part 73 RIN 2120-AA66 Amendment of Multiple Restricted Areas;...

  2. Pilot-scale in situ vitrification at Arnold Engineering Development Center, Arnold AFB, TN

    SciTech Connect

    Lominac, J.K.; Edwards, R.C. ); Timmerman, C.L. )

    1989-11-01

    The Department of Defense has the Installation Restoration Program (IRP) to identify and permanently remediate hazardous material disposal sites at its military bases across the United States. Pursuant to this guidance, Arnold Engineering Development Center (AEDC) selected In Situ Vitrification (ISV) to remediate an old fire training area, Fire Protection Training Area (FPTA) No. 2. The ISV technology was developed by Pacific Northwest Laboratory, Richland, WA, for the US Department of Energy (DOE) and will allow for the destruction and encapsulation of the petroleum-oil-lubricants (POL) and heavy metal-constituents found at the FPTA and adjacent overflow pond. ISV operates by passing a measured current of electricity into the ground through a set of electrodes. The resulting heat causes the soil to melt and form a solid vitreous (glass) mass similar to naturally occurring obsidian or basalt. In the process, organic constituents will be pyrolyzed (changed by heat) by the ensuing heat whereas the non-organic material will be incorporated into the glass matrix. 2 refs., 9 figs.

  3. OPTIMIZATION OF IN-SITU THERMAL REMEDIATION: THE LORING AFB STEAM INJECTION PROJECT EXAMPLE

    EPA Science Inventory

    Environmental remediation programs require that adequate planning be done before field work for characterization or remediation is undertaken. However, the heterogeneous nature of the subsurface can often thwart our best planning efforts. More recently, dynamic work plans which...

  4. Experimental plan and construction guidance for Hanford Protective Barrier Test at Hill AFB, Utah

    SciTech Connect

    Kirkham, R.R.; Gee, G.W.

    1994-04-01

    Permanent isolation surface barriers are needed to fully isolate buried waste materials for long periods of time from the general environment. Multi-layer surface barriers that use natural earthen materials overlying bio-intrusion layers of rock and/or asphalt are expected to reduce infiltration or deep percolation to less than 0.5 mm (0.02 in.) of water. The layering also reduces root penetration, preventing uptake and translocation of hazardous materials to the surface vegetation. The opportunity exists to test the Hanford Barrier in a wetter, colder climate at an existing lysimeter facility located at Hill Air Force Base near Salt Lake, Utah. The use of an existing lysimeter facility simplifies the construction effort and allows comparison of the Hanford Protective Barrier with an existing US Environmental Protection Agency-Resource Conservation and Recovery Act clay cap already undergoing tests at the lysimeter site. This document presents the experimental plan for testing the Hanford Protective Barrier in a wetter, colder climate.

  5. Views of STS-4 crew and families at Ellington AFB departing for KSC

    NASA Technical Reports Server (NTRS)

    1982-01-01

    Views of STS-4 crew and families at Ellington Air Force Base departing for Kennedy Space Center. Astronauts Thomas K. Mattingly, II., right, and Henry W. Hartsfield, Jr. on the Ellington Air Force Base runway. Behind them are the T-38's they will be flying to the Kennedy Space Center (32917,32924); Mattingly climbs aboard NASA aircraft 918, a T-38 jet, as he prepares to leave Ellington for KSC (32918); Mattingly seating in T-38 with canopy up preparing to leave (32919,32925); Hartsfield prepares to board NASA aircraft 919, a T-38 jet, at Ellington (32920); Hartsfield seated in T-38 with canopy up preparing to leave Ellington (32921,32923); The family of Astronuat Henry W. Hartsfield, Jr. stands near the NASA Gulfstream aircraft priot to boarding it for Florida. Astronaut Bryan D. O'Conner talks with the family - from the left, daughters Judy Lynn, Keely Warren and wife Judy Frances Hartsfield (32922).

  6. Astronauts McDivitt and White during water egress training at Ellington AFB

    NASA Technical Reports Server (NTRS)

    1965-01-01

    Astronauts James A. McDivitt, command pilot, and Edward H. White II, pilot, the Gemini-Titan 4 prime crew are seen during water egress training at Ellington Air Force Base, Texas. Views include McDivitt climbing out of the capsule and into the water beside White while a diver looks on. Both are wearing full space suits (19282); White floats in a life raft while McDivitt clings to a life vest in the pool with the capsule floating nearby (19283).

  7. NOAA-L satellite is mated to Apogee Kick Motor at Vandenberg AFB

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Inside the B16-10 spacecraft processing hangar at Vandenberg Air Force Base, Calif., workers oversee the mating of the Apogee Kick Motor (below) to the National Oceanic and Atmospheric Administration (NOAA-L) satellite above. NOAA-L is part of the Polar-Orbiting Operational Environmental Satellite (POES) program that provides atmospheric measurements of temperature, humidity, ozone and cloud images, tracking weather patterns that affect the global weather and climate. The launch of the NOAA-L satellite is scheduled no earlier than Sept. 12 aboard a Lockheed Martin Titan II rocket. Tuberculosis in Australia: bacteriologically confirmed cases and drug resistance, 2000: report of the Australian Mycobacterium Laboratory Reference Network.

    PubMed

    Lumb, Richard; Bastian, Ivan; Dawson, David; Gilpin, Chris; Havekort, Frank; Howard, Peter; Sievers, Aina

    2002-01-01

    The Australian Mycobacterium Reference Laboratory Network collected and analysed laboratory data on new diagnoses of disease caused by Mycobacterium tuberculosis complex in the year 2000. A total of 765 cases were identified, representing an annual reporting rate of 4.0 cases of laboratory-confirmed tuberculosis (TB) per 100,000 population. Pulmonary disease was diagnosed in 64.9 per cent of cases with a male:female ratio of 1.5:1. Smears were positive for 209/365 (57.3%) of sputum isolates and 39/117 (33.3%) bronchoscopy isolates. Sputum from males was more likely to be smear-positive (63.3%) than from females (47.5%). Isolates from lymph node accounted for 136 (17.7%) of all cases; only 28.7 per cent were smear-positive. Eighty-four (11.0%) isolates, comprising 82 M. tuberculosis and 2 M. bovis strains, demonstrated in vitro resistance to at least one of the standard anti-TB medications. Resistance to at least isoniazid and rifampicin (defined as multidrug-resistant TB) was observed for only 8 (1.0%) strains, a rate similar to previous years. Almost all (96.3%) of patients with drug resistant strains were classified as having initial resistance. The country of birth was known for 76 (92.7%) of 82 patients with a drug resistant strain of M. tuberculosis; 6 were Australian-born and 70 (92.1%) had migrated from a total of 17 countries. Of these 70 migrants with drug-resistant disease, 68.6 per cent had migrated from one of the following countries: Vietnam (n=15), China (n=11), Philippines (n=11), India (n=6), and Indonesia (n=5).

  8. Tuberculosis in Australia: bacteriologically confirmed cases and drug resistance, 2000: report of the Australian Mycobacterium Laboratory Reference Network.

    PubMed

    Lumb, Richard; Bastian, Ivan; Dawson, David; Gilpin, Chris; Havekort, Frank; Howard, Peter; Sievers, Aina

    2002-01-01

    The Australian Mycobacterium Reference Laboratory Network collected and analysed laboratory data on new diagnoses of disease caused by Mycobacterium tuberculosis complex in the year 2000. A total of 765 cases were identified, representing an annual reporting rate of 4.0 cases of laboratory-confirmed tuberculosis (TB) per 100,000 population. Pulmonary disease was diagnosed in 64.9 per cent of cases with a male:female ratio of 1.5:1. Smears were positive for 209/365 (57.3%) of sputum isolates and 39/117 (33.3%) bronchoscopy isolates. Sputum from males was more likely to be smear-positive (63.3%) than from females (47.5%). Isolates from lymph node accounted for 136 (17.7%) of all cases; only 28.7 per cent were smear-positive. Eighty-four (11.0%) isolates, comprising 82 M. tuberculosis and 2 M. bovis strains, demonstrated in vitro resistance to at least one of the standard anti-TB medications. Resistance to at least isoniazid and rifampicin (defined as multidrug-resistant TB) was observed for only 8 (1.0%) strains, a rate similar to previous years. Almost all (96.3%) of patients with drug resistant strains were classified as having initial resistance. The country of birth was known for 76 (92.7%) of 82 patients with a drug resistant strain of M. tuberculosis; 6 were Australian-born and 70 (92.1%) had migrated from a total of 17 countries. Of these 70 migrants with drug-resistant disease, 68.6 per cent had migrated from one of the following countries: Vietnam (n=15), China (n=11), Philippines (n=11), India (n=6), and Indonesia (n=5). PMID:12206373

  9. Major Delays in the Diagnosis and Management of Tuberculosis Patients in Nepal

    PubMed Central

    Mahato, Roshan Kumar; Vaeteewootacharn, Kriangsak; Koju, Rajendra; Bhattarai, Ratna

    2015-01-01

    Background Early diagnosis is a determining factor for spread of tuberculosis. Delay in diagnosis and treatment of tuberculosis geometrically increases spread and infectivity of the disease and is associated with higher risk of mortality. Aim The present study aimed to investigate the length of delays in diagnosis and treatment among new pulmonary tuberculosis patients in central development region of Nepal. Materials and Methods A cross-sectional study was conducted by administration of structured questionnaire interview and reviewing the medical records of the new sputum smear positive pulmonary tuberculosis cases during January–May 2015. Simple random sampling was applied to select samples from 5 districts of 19 districts comprising at least one each from 3 ecological regions of Nepal. Results A total of 374 new sputum smear positive pulmonary tuberculosis cases were included in the study. The median patient delay, health system delay, and total delay were 32 days, 3 days and 39.5 days respectively. The unacceptable patients delay was 53.21% (95% CI: 48.12-58.29) of all new patients, whereas it was 26.74% (95% CI: 22.23-31.24) for the unacceptable health system delay and the unacceptable total delay was 62.83% (95% CI: 57.91-67.75). Conclusion TB diagnosis and treatment is still a significant problem of Nepal. Majority of unacceptable delays were from patients. Identifying factors influencing delays and developing evidence-based approaches to address those delays will help in advancing tuberculosis prevention and management in low-income settings. PMID:26557545

  10. Use of GeneXpert Mycobacterium tuberculosis/rifampicin for rapid detection of rifampicin resistant Mycobacterium tuberculosis strains of clinically suspected multi-drug resistance tuberculosis cases

    PubMed Central

    Guenaoui, Kheira; Ouardi, Aissa; Zeggai, Soumia; Sellam, Feriel; Bekri, Farid; Cherif Touil, Sakina

    2016-01-01

    Background Multi-drug resistance (MDR) TB is defined as tuberculosis (TB) disease caused by a strain of Mycobacterium tuberculosis (MTB) that was resistant to at least isoniazid and rifampicin (RIF). Emerging Multidrug-Resistant TB is one of the major concerns of health policy and rapid detection of M. tuberculosis and detection of RIF resistance in infected patients are essential for disease management. The aim of this study was to evaluate patterns of RIF resistance in cases of sputum positive pulmonary TB by using GeneXpert MTB/RIF and comparing between phenotypic and genotypic testing of RIF resistance in MTB strains of clinically suspected MDR-TB isolated cases in western Algeria. Methods In this study 50 sputum positive cases of pulmonary TB who were potential MDR suspect were included. Their sputum samples were collected and subjected to sputum smear microscopy, culture and conventional MTB/RIF test followed by GeneXpert MTB/RIF assay. Results Of total 50 cases included in this study, MTB was detected in all patients (100%) by GeneXpert MTB/RIF. However, RIF’s resistance was detected in only 21 cases (42%) by GeneXpert MTB/RIF. All RIF resistant strains detected by GeneXpert MTB/RIF were phenotypically confirmed as MDR strains. 42.85% of cases were retreatment failure cases, retreatment cases smear positive at 4 months were 23.82%. While 19.05% of cases were retreatment cases smear positive at diagnosis, and 14.28% patient had history of contact with MDR-TB. Sensitivity, specificity, positive predictive value and negative predictive value of Xpert MTB/RIF to detect RIF resistance in comparison to conventional phenotypic drug susceptibility technique were found equal to the rates of 100%, 100%, 100% and 100%, respectively. Conclusions GeneXpert MTB/RIF assay is efficient and reliable technique for the rapid diagnostic of TB. It’s simplicity, high sensitivity and specificity for RIF resistance detection make this technique a very attractive tool for

  11. Multicenter evaluation of a real-time loop-mediated isothermal amplification (RealAmp) test for rapid diagnosis of Mycobacterium tuberculosis.

    PubMed

    Ou, Xichao; Wang, Shengfen; Dong, Haiyan; Pang, Yu; Li, Qiang; Xia, Hui; Qu, Yan; Zhang, Zhiying; Li, Junchen; Zhang, Jack; Huan, Shitong; Chin, Daniel P; Zhao, Yanlin

    2016-10-01

    The real-time loop-mediated isothermal amplification (RealAmp) test is a novel technology for rapid diagnosis of Mycobacterium tuberculosis (MTB) from sputum samples. This test has not previously been effectively evaluated. In this study, we assessed the performance of the RealAmp test for tuberculosis (TB) detection among TB suspected patients from three provincial hospitals in China. Each eligible TB suspected patients in Shanghai, Guangzhou, and Shandong provided one sputum specimen each, smear microscopy, liquid culture, and the RealAmp test were performed per patient. A total of 1583 TB suspected patients were enrolled in the study, and 1519 were used for the performance analysis. The sensitivity and specificity of the RealAmp test for MTB detection were calculated using liquid culture as reference method. The sensitivity of RealAmp test in smear-negative and culture-positive patients was 60.08%; the sensitivity in smear-positive and culture-positive patients was 97.09%; and the overall sensitivity in culture-positive patients was 74.88%. The specificity of RealAmp test for MTB detection was 86.50%.

  12. Rapid diagnosis of MDR and XDR tuberculosis with the MeltPro TB assay in China.

    PubMed

    Pang, Yu; Dong, Haiyan; Tan, Yaoju; Deng, Yunfeng; Cai, Xingshan; Jing, Hui; Xia, Hui; Li, Qiang; Ou, Xichao; Su, Biyi; Li, Xuezheng; Zhang, Zhiying; Li, Junchen; Zhang, Jiankang; Huan, Shitong; Zhao, Yanlin

    2016-01-01

    New diagnostic methods have provided a promising solution for rapid and reliable detection of drug-resistant TB strains. The aim of this study was to evaluate the performance of the MeltPro TB assay in identifying multidrug-resistant (MDR-) and extensively drug-resistant tuberculosis (XDR-TB) patients from sputum samples. The MeltPro TB assay was evaluated using sputum samples from 2057 smear-positive TB patients. Phenotypic Mycobacterial Growth Indicator Tube (MGIT) 960 drug susceptibility testing served as a reference standard. The sensitivity of the MeltPro TB assay was 94.2% for detecting resistance to rifampicin and 84.9% for detecting resistance to isoniazid. For second-line drugs, the assay showed a sensitivity of 83.3% for ofloxacin resistance, 75.0% for amikacin resistance, and 63.5% for kanamycin resistance. However, there was a significant difference for detecting kanamycin resistance between the two pilot sites in sensitivity, which was 53.2% in Guangdong and 81.5% in Shandong (P = 0.015). Overall, the MeltPro TB assay demonstrated good performance for the detection of MDR- and XDR-TB, with a sensitivity of 86.7% and 71.4%, respectively. The MeltPro TB assay is an excellent alternative for the detection of MDR- and XDR-TB cases in China, with high accuracy, short testing turn-around time, and low unit price compared with other tests. PMID:27149911

  13. Multicenter evaluation of a real-time loop-mediated isothermal amplification (RealAmp) test for rapid diagnosis of Mycobacterium tuberculosis.

    PubMed

    Ou, Xichao; Wang, Shengfen; Dong, Haiyan; Pang, Yu; Li, Qiang; Xia, Hui; Qu, Yan; Zhang, Zhiying; Li, Junchen; Zhang, Jack; Huan, Shitong; Chin, Daniel P; Zhao, Yanlin

    2016-10-01

    The real-time loop-mediated isothermal amplification (RealAmp) test is a novel technology for rapid diagnosis of Mycobacterium tuberculosis (MTB) from sputum samples. This test has not previously been effectively evaluated. In this study, we assessed the performance of the RealAmp test for tuberculosis (TB) detection among TB suspected patients from three provincial hospitals in China. Each eligible TB suspected patients in Shanghai, Guangzhou, and Shandong provided one sputum specimen each, smear microscopy, liquid culture, and the RealAmp test were performed per patient. A total of 1583 TB suspected patients were enrolled in the study, and 1519 were used for the performance analysis. The sensitivity and specificity of the RealAmp test for MTB detection were calculated using liquid culture as reference method. The sensitivity of RealAmp test in smear-negative and culture-positive patients was 60.08%; the sensitivity in smear-positive and culture-positive patients was 97.09%; and the overall sensitivity in culture-positive patients was 74.88%. The specificity of RealAmp test for MTB detection was 86.50%. PMID:27425377

  14. Treatment Outcomes of Patients Placed on Treatment Under Directly Observed Therapy Short-Course (Dots)

    PubMed Central

    Kaur, Gurpreet; Goel, N.K.; Kumar, Dinesh; Janmeja, A.K.; Swami, H.M.; Kalia, Meenu

    2008-01-01

    Background: Tuberculosis continues to be a pressing health problem in India. The Revised National Tuberculosis Programme (RNTCP), an application of Directly Observed Treatment Short-course (DOTS) in India, launched in 1997 needs continuous evaluation. Objective: To study the outcomes of treatment among the patients put on DOTS under RNTCP in Chandigarh, UT. Material & Methods: A Longitudinal study was conducted during 2004-2005 in 13 Microscopic centres (MC's) spread over 2 Tuberculosis Units (TU's) under District Tuberculosis Centre (DTC) in Union Territory (UT), Chandigarh. A sample of 265 respondents, selected by two-stage stratified random sampling technique, was recruited in the study cohort. Data analysis was done using SPSS-10 statistical software package. Results: For Category I and Category II patients, the Success rate was 98.6% and 90.4% respectively. The overall default rate was 1.1% and failure rate was 2.6%. For re-treatment cases, failure rate was higher i.e. 5.8%. The sputum conversion rate among the new smear positive cases was 93.8% at 3 months of treatment. For the re-treatment cases, spu-tum conversion rate at 3 months was 94.1%. Conclusion: The study concludes that RNTCP is running successfully in UT Chandigarh, having high success rate and low default rate. The reasons for high failure rate should be explored in depth. PMID:20165654

  15. Rapid diagnosis of MDR and XDR tuberculosis with the MeltPro TB assay in China

    PubMed Central

    Pang, Yu; Dong, Haiyan; Tan, Yaoju; Deng, Yunfeng; Cai, Xingshan; Jing, Hui; Xia, Hui; Li, Qiang; Ou, Xichao; Su, Biyi; Li, Xuezheng; Zhang, Zhiying; Li, Junchen; Zhang, Jiankang; Huan, Shitong; Zhao, Yanlin

    2016-01-01

    New diagnostic methods have provided a promising solution for rapid and reliable detection of drug-resistant TB strains. The aim of this study was to evaluate the performance of the MeltPro TB assay in identifying multidrug-resistant (MDR-) and extensively drug-resistant tuberculosis (XDR-TB) patients from sputum samples. The MeltPro TB assay was evaluated using sputum samples from 2057 smear-positive TB patients. Phenotypic Mycobacterial Growth Indicator Tube (MGIT) 960 drug susceptibility testing served as a reference standard. The sensitivity of the MeltPro TB assay was 94.2% for detecting resistance to rifampicin and 84.9% for detecting resistance to isoniazid. For second-line drugs, the assay showed a sensitivity of 83.3% for ofloxacin resistance, 75.0% for amikacin resistance, and 63.5% for kanamycin resistance. However, there was a significant difference for detecting kanamycin resistance between the two pilot sites in sensitivity, which was 53.2% in Guangdong and 81.5% in Shandong (P = 0.015). Overall, the MeltPro TB assay demonstrated good performance for the detection of MDR- and XDR-TB, with a sensitivity of 86.7% and 71.4%, respectively. The MeltPro TB assay is an excellent alternative for the detection of MDR- and XDR-TB cases in China, with high accuracy, short testing turn-around time, and low unit price compared with other tests. PMID:27149911

  16. Identification of multidrug resistance in previously treated tuberculosis patients: a mixed methods study in Cambodia

    PubMed Central

    Royce, S; Khann, S; Yadav, RP; Mao, ET; Cattamanchi, A; Sam, S; Handley, MA

    2014-01-01

    SUMMARY Setting Previously treated tuberculosis (TB) patients are a priority for drug susceptibility testing (DST) to identify cases with multidrug resistance (MDR). In Cambodia, a recent study found that only one-third of smear-positive previously treated patients had DST results. Objective To quantify the gaps in detecting MDR in previously treated TB patients in Cambodia, and describe health workers’ perspectives on barriers, facilitators and potential interventions. Design We analyzed case notifications in Cambodia (2004–2012) and conducted semi-structured interviews with key stakeholders Results The proportion of previously treated notifications varied significantly across provinces 2010–12, in the context of longer term trends of decreasing relapse and increasing “other” retreatment notifications. Correct classification of patients’ TB treatment history and ensuring specimens from previously-treated patients are collected and reach the laboratory could nearly double the number of detected MDR-TB cases. Identified barriers include patients’ reluctance to disclose and staff difficulty eliciting treatment history, partly due to availability of streptomycin only in hospitals. Facilitators include trained health workers, collection of sputum for DST even if previously treated patients are not taking streptomycin, streamlining sputum transportation and promptly reporting results. Conclusion Improved monitoring, supportive supervision, and correctly classifying previously treated patients are essential for improving detection of MDR-TB. PMID:25299861

  17. Randomized, placebo-controlled phase II trial of heat-killed Mycobacterium vaccae (Longcom batch) formulated as an oral pill (V7)

    PubMed Central

    Efremenko, Yuri V; Butov, Dmytro A; Prihoda, Natalia D; Zaitzeva, Svetlana I; Yurchenko, Larisa V; Sokolenko, Nina I; Butova, Tetyana S; Stepanenko, Anna L; Kutsyna, Galyna A; Jirathitikal, Vichai; Bourinbaiar, Aldar S

    2013-01-01

    One-month Phase II trial was conducted in 43 sputum smear-positive patients with pulmonary tuberculosis randomized into treatment (n = 22) and placebo (n = 21) arms to investigate the safety and efficacy of an orally-administered therapeutic TB vaccine (V7) containing 10 μg of heat-killed Mycobacterium vaccae provided by Longcom company. Immunotherapy and control groups comprised 8 newly diagnosed (1stDx TB; 18.6%), 6 re-treated (RTB; 14%), and 29 multidrug-resistant (MDR-TB; 67.4%) cases distributed at 5:4:13 and 3:2:16 ratios, respectively. Both arms received conventional TB drugs administered under directly observed therapy. The average weight gain in V7 arm was modest, but statistically significant (0.6 kg; p = 0.004), while placebo patients lost 0.1 kg (p = 0.77). Except defervescence and increased lymphocyte percentage, other secondary endpoints such as erythrocyte sedimentation rate (ESR), leukocyte counts and hemoglobin content were not significantly affected. In control patients only one secondary endpoint, ESR, has improved. After one month mycobacterial clearance in sputum smears was observed in 31.8% (p = 0.03) and 9.5% (p = 0.83) of patients on V7 and placebo. However, the difference between outcomes in two arms was below significance threshold (p = 0.07). Thus, larger population of patients with prolonged follow-up is required to support these preliminary findings. PMID:23782489

  18. [Pulmonary infection caused by Mycobacterium gordonae (M. gordonae) in a healthy middle-aged male].

    PubMed

    Hasegawa, T; Tada, K; Ishii, M

    1992-02-01

    A 51-year-old man was admitted to our hospital in July 1989 because of an abnormality in his chest radiograph. On his yearly health check-up, an abnormality of his chest radiography was first noted in June 1988. At that time, examinations including bronchoscopy were performed but no specific diagnosis was made. On admission, his chest radiograph revealed new infiltrates at the apex of the right lung which were not present in June 1988. Three out of 5 consecutive sputum specimens after admission produced a pure growth of 100 colonies to 1+ of acid-fast bacilli (AFB). This AFB was scotochromogenic, and hydrolysis of Tween 80 at 5 days was positive. It did not reduce nitrate, and niacin test was negative. It was sensitive to ethambutol at a concentration of 5 micrograms/ml, and was not tolerant to 0.2% picric acid. We thus identified this AFB to be M. gordonae. The patient was treated with rifampicin (450 mg/day), isoniazid (400 mg/day), and ethambutol (1000 mg/day) for 9 months. After 2 months of treatment the sputum cultures became negative, and the chest radiograph showed improvement of the infiltrates. M. gordonae is considered to be one of the least pathogenic AFB to man. Most recent reports of M. gordonae infection have been in immunocompromized hosts or patients with a history of pulmonary tuberculosis. The present case is a very rare example of this organism affecting a healthy male, and thus yields new information on the pathogenesis of M. gordonae in man.

  19. [Tuberculosis in Asia].

    PubMed

    2002-10-01

    assistance of Medical Committee Netherlands Vietnam (MCNV). The New National TB Control Program follows the main directives of WHO and IUATLD's procedures of case-finding, chemotherapy and management. Passive case-findings are based on sputum smear. Chemotherapy with priority for smear positive cases is 3SHZ/6S2H2 for new cases and 3HRE/6H2R2E2 for retreated cases, which is undertaken with directly observed therapy (DOT strategy) mainly at commune health posts. Since 1989, DOTS strategy with 2SHRZ/6HE for new cases and 2SHRZE/1HRZE/5H3R3E3 for retreated cases has gradually been introduced in districts and communes of every province. In 1995, the government established the National and Provincial TB Control Steering Committees and has provided incentives for detected smear positive cases and cured smear positive cases. The government has also started strengthening the program of managerial and supervisory capacity for TB staff and has promoted the cooperation of all associated organizations of TB control. The WHO global surveillance and monitoring project reports that in 2000 Vietnam reached the global target, i.e., 99.8% population covered by DOTS with 80% of expected new smear positive cases being detected and a high cure rate ranging from 85.3% in 1989 to 90.3% in 1999. A distinguishing aspect of TB control in Vietnam is the effective international partnerships combined with high political commitment of the government nationally and provincially as well as active participation of all organizations in the community.