Correlation between the shape of the ion mobility signals and the stepwise folding process of polylactide ions.

In the field of polymer characterization, the use of ion mobility mass spectrometry (IMMS) remains mainly devoted to the temporal separation of cationized oligomers according to their charge states, molecular masses and macromolecular architectures in order to probe the presence of different structures. When analyzing multiply charged polymer ions by IMMS, the most striking feature is the observation of breaking points in the evolution of the average collision cross sections with the number of monomer units. Those breaking points are associated to the folding of the polymer chain around the cationizing agents. Here, we scrutinize the shape of the arrival time distribution (ATD) of polylactide ions and associate the broadening as well as the loss of symmetry of the ATD signals to the coexistence of different populations of ions attributed to the transition from opened to folded stable structures. The observation of distinct distributions reveals the absence of folded/extended structure interconversion on the ion mobility time scale (1-10 ms) and then on the lifetime of ions within the mass spectrometer at room temperature. In order to obtain information on the possible interconversion between the different observed populations upon ion activation, we performed IM-IM-MS experiments (tandem ion mobility measurements). To do so, mobility-selected ions were activated by collisions before a second mobility measurement. Interestingly, the conversion by collisional activation from a globular structure into a (partially) extended structure, i.e. the gas phase unfolding of the ions, was not observed in the energetic regime available with the used experimental setup. The absence of folded/extended interconversion, even upon collisional activation, points to the fact that the polylactide ions are 'frozen' in their specific 3D structure during the desolvation/ionization electrospray processes. Copyright © 2017 John Wiley & Sons, Ltd.

GenoType® MTBDRplus assay detection of drug-resistant tuberculosis in routine practice in Korea.

SETTING: Korea is an intermediate-burden country with high rates of tuberculosis (TB) drug resistance. OBJECTIVE: To evaluate the performance of the GenoType® MTBDRplus (MTBDR) assay in diagnosing drug-resistant TB in routine practice in Korea. DESIGN: The MTBDR assay was performed on 428 samples, and the results were retrospectively compared with the results of conventional drug susceptibility testing (DST). The interval between treatment and diagnosis of drug resistance was also compared. RESULTS: The sensitivity, specificity and positive and negative predictive values of the MTBDR assay were respectively 96.6%, 98.9%, 93.4% and 99.5% for the detection of rifampicin (RMP) resistance; 93.8%, 98.3%, 92.7% and 98.6% for isoniazid (INH) resistance; and 91.1%, 99.2%, 99.4% and 98.7% for multidrug-resistant TB (MDR-TB). The median interval between the start of anti-tuberculosis chemotherapy and the reporting of results was 88.9 days for conventional DST and 19.8 days for MTBDR using clinical specimens. CONCLUSION: The specificity of the MTBDR assay in detecting MDR-TB was very high, although the sensitivity in detecting INH resistance and MDR-TB was not optimal (<95%). Although the turnaround time in detecting drug resistance was dramatically reduced with MTBDR compared to conventional DST, more effort is needed to shorten the turnaround time.

Proteins involved in DNA damage response pathways and survival of stage I non-small-cell lung cancer patients.

BACKGROUND: Biological complexity leads to significant variation in the survival of patients with stage I non-small-cell lung cancer (NSCLC). DNA damage response (DDR) pathways play a critical role in maintaining genomic stability and in the progression of NSCLC. Therefore, the development of a prognostic biomarker focusing on DDR pathways is an intriguing issue. PATIENTS AND METHODS: Expression of several proteins (ATM, ATMpS1981, γH2AX, 53BP1, 53BP1pS25, Chk2, Chk2pT68, MDC1, MDC1pS964, BRCA1pS1423, and ERCC1) and overall survival were investigated in 889 pathological stage I NSCLC patients. RESULTS: Low expression of BRCA1pS1423 or ERCC1 was significantly associated with worse survival in the whole cohort of patients. Analysis performed based on histology revealed that low expression of γH2AX, Chk2pT68, or ERCC1 was a poor prognostic factor in squamous cell carcinoma patients [adjusted hazard ratio (aHR), Cox P: 1.544, 0.012 for γH2AX; 1.624, 0.010 for Chk2pT68; 1.569, 0.011 for ERCC1]. The analysis of the interaction between two proteins showed that this effect was more pronounced in squamous cell carcinoma patients. However, these effects were not detected in adenocarcinoma patients. CONCLUSIONS: The proteins involved in DDR pathways exhibited differential expression between squamous cell carcinoma and adenocarcinoma and were important determinants of survival in stage I squamous cell carcinoma patients.

Distinguishing tuberculosis from Mycobacterium avium complex disease using an interferon-gamma release assay.

SETTING: The QuantiFERON-TB Gold (QFT-G) test can be used to distinguish between tuberculosis (TB) and non-tuberculous mycobacterial disease, but a high background TB infection rate may pose a problem. Although the QuantiFERON-TB (QFT) test, the first-generation QFT-G test, employs a non-specific PPD antigen, avium sensitin is also used as a stimulating antigen. OBJECTIVE: To evaluate the utility of these two interferon gamma release assays (IGRAs), QFT-G and QFT, and the tuberculin skin test (TST), to differentiate TB from Mycobacterium avium complex (MAC) disease in an intermediate TB burden country. METHODS: We compared the diagnostic performance of these three tests in 38 prospectively enrolled patients with TB and 40 with MAC lung disease. RESULTS: The TST yielded positive results in 70.6% of TB and 47.5% of MAC patients; the proportions were respectively 89.5% and 34.3% for QFT-G and 86.8% and 35.3% for QFT. The three tests were of similar accuracy, sensitivity and specificity in diagnosing TB. CONCLUSION: Our findings indicate that the TST and IGRAs could not discriminate between active TB and MAC disease or latent TB infection in a TB-endemic area.

Non-tuberculous mycobacterial diseases presenting as solitary pulmonary nodules.

SETTING: South Korea, 2005-2007. BACKGROUND: Non-tuberculous mycobacterial diseases presenting as solitary pulmonary nodules (NTM-SPN) are rare and may be misdiagnosed as tuberculoma. DESIGN: Eleven NTM-SPN patients were enrolled retrospectively and compared with two groups each of 33 patients with upper lobe cavitary (UC) and nodular bronchiectatic (NB) type NTM diseases, respectively. RESULTS: The patients' median age was 52 years. Compared with UC-type disease, NTM-SPN patients showed female (n = 9) and never smoker (n = 8) predominance and fewer comorbidities (n = 0; P < 0.05 for each). Mycobacterium avium (n = 9) was predominant in NTM-SPN patients, followed by M. intracellulare (n = 2), whereas M. intracellulare was most frequently isolated from patients with both UC- and NB-type NTM (P < 0.05). The nodules were a median of 2.3 cm in diameter (range 1.6-6.3 cm) and were distributed evenly in all lobes. In all patients, percutaneous needle aspiration/biopsy was performed with a median 71 days of diagnostic delay. Ten patients successfully completed treatment, and one showed spontaneous reduction of nodule size without treatment. Reverse blot hybridisation assays of six DNA samples identified four subjects with M.avium-intracellulare complex (MAC), in line with conventional test data. CONCLUSION: NTM-SPN was caused exclusively by MAC. Although clinical outcome was favourable, confirmatory diagnosis was delayed. Molecular methods are needed for early diagnosis of NTM-SPN.

Incidence of tuberculosis among health care workers at a private university hospital in South Korea.

SETTING: A private university hospital in South Korea. OBJECTIVE: To investigate the incidence of tuberculosis (TB) among health care workers (HCWs) employed at a university hospital in South Korea. DESIGN: The occurrence of TB cases among HCWs over a 6-year period (2001-2006) was reviewed. The prevalence of TB was compared with that of the general population using a prevalence ratio (PR) adjusted by age and sex. RESULTS: The study involved 8433 people. TB developed in 61 HCWs (0.72%). The prevalence ratio among all HCWs was 1.05 (95%CI 0.80-1.35). In occupational subgroups, the PRs for doctors, nurses and employees in other departments were respectively 0.58 (95%CI 0.30-1.01), 1.81 (95%CI 1.21-2.59) and 0.95 (95% CI 0.58-1.46). For nurses working in TB-related departments, the relative risk of developing TB was 3.4 times higher (95%CI 1.52-8.25) than for employees in other departments (P=0.005). The PR for nurses working in TB-related department was 5.1 (95%CI 3.23-8.42). CONCLUSION: Among HCWs, nurses working in TB-related departments were at increased risk of developing TB. This suggests that some TB develops via in-hospital infection.

High isolation rate of adenovirus serotype 7 from South Korean military recruits with mild acute respiratory disease.

Adenovirus is a major cause of acute respiratory disease (ARD) in military recruits. When South Korean military recruits with ARD were surveyed, adenovirus was identified in 122 (61.0%) of the 200 recruits studied. Moreover, all cases of ARD involving adenovirus were caused by serotype 7.

Role of the C-reactive protein for the diagnosis of TB among military personnel in South Korea.

Clinicians are frequently faced with the task of differentiating between pulmonary tuberculosis (PTB) and pneumonia. We evaluated the role of the C-reactive protein test (CRP) for differentiating between TB and pneumonia among military personnel in South Korea. Only immunocompetent males were eligible. Forty-six patients with PTB and 67 with pneumonia were enrolled prospectively. Median CRP concentration was lower in patients with TB than in patients with non-tuberculous pneumonia (3.2 mg/dl [range 0.1-15.7 mg/dl] vs. 8.3 mg/dl [range 0.2-33.7 mg/dl], P < 0.001). The sensitivity and specificity for TB of a low CRP concentration (< 11.2 mg/dl) in serum was 93.3% and 40.9%, respectively. CRP concentration measurement might be useful for eliminating the diagnosis of TB.

The role of TST in the diagnosis of latent tuberculosis infection among military personnel in South Korea.

BACKGROUND: The rapid and accurate diagnosis of latent tuberculosis infection (LTBI) is crucial in military settings because military personnel live in crowded circumstances and are of an age group with a high incidence of tuberculosis (TB). We tried to elucidate whether the tuberculin skin test (TST) accurately reflects the risk of TB infection among military personnel, in a setting of intermediate TB prevalence and where bacille Calmette-Guérin (BCG) vaccination is mandatory. METHODS: A multi-stage cluster survey was conducted among military personnel in South Korea. Participants were grouped according to their risk of TB infection: Group 1, no identifiable risk of TB; Groups 2 and 3, recent casual (Group 2) or close (Group 3) contact with smear-positive TB patients. RESULTS: Of 1045 participants, 857 (82.0%) had been BCG-vaccinated. The odds ratio (OR) of a positive TST (10 mm cut-off) for Group 2 (n = 184) and Group 3 (n = 83) compared with Group 1 (n = 778) were 0.95 (95%CI 0.67-1.38) and 1.7 (95%CI 1.06-2.70), respectively (P value for trend 0.16). CONCLUSIONS: The TST does not accurately reflect the risk of LTBI among young military personnel in a setting where there is intermediate TB prevalence and extensive BCG coverage.

Pulmonary tuberculosis in young Korean soldiers: incidence, drug resistance and treatment outcomes.

OBJECTIVE: To determine the incidence and treatment outcomes of pulmonary tuberculosis (PTB) in young soldiers of South Korea. DESIGN: From 2000 to 2004, all soldiers with a new diagnosis of tuberculosis (TB) were enrolled in the study, based on the official records of the Armed Forces Medical Command. The demographic and clinical data of the cases were evaluated retrospectively. RESULTS: A total of 3115 TB cases were reported during the study period, of whom 2071 (66.5%) were reported as PTB. The annual incidence rates of PTB were 96.4 per 100,000 population in 2000, 89.3 in 2001, 67.6 in 2002, 60.2 in 2003, and 63.1 in 2004. A total of 270 patients diagnosed and treated at the Armed Forces Capital Hospital were analysed. Of the Mycobacterium tuberculosis isolates, 87.4% were susceptible to all available anti-tuberculosis drugs; 253 (93.7%) patients eventually completed initial anti-tuberculosis treatment. Among the patients with smear-positive PTB, the cure rate was 89.3% (100/112). CONCLUSION: Our results demonstrate that the incidence of PTB in Korean soldiers, although still high, was declining steadily. With good case management, the overall success rate of initial treatment was approximately 90%.

Oral insertion of a flexible bronchoscope is associated with less discomfort than nasal insertion for Korean patients.

OBJECTIVE: The route of bronchoscope insertion varies between centres, without a firm rationale based on well-designed studies. We therefore compared nasal and oral insertion of a flexible bronchoscope and evaluated efficacy and patient satisfaction. DESIGN: Prospective randomised study of patients who underwent flexible bronchoscopy from May to September 2003 and who were randomly assigned to nasal and oral insertion approaches. RESULTS: Clinical characteristics, factors related to the procedure and patient satisfaction were analysed. In total, 307 patients were randomly assigned to the nasal (n = 158) or oral insertion groups (n = 149). No difference in baseline characteristics was identified between the groups. Insertion by the oral route was associated with a smaller amount of lidocaine use during the procedure (P = 0.04) and less frequent insertion site bleeding (P = 0.005). Patients assigned to oral insertion reported less discomfort during anaesthesia (P = 0.01) and scope insertion (P < 0.001), as well as less dyspnoea (P = 0.04) and coughing (P = 0.03). CONCLUSION: Oral insertion of a flexible bronchoscope was associated with less discomfort for patients than nasal insertion, although the route of insertion had no significant effect on outcome.

Leishmaniasis as an emerging infection.

Leishmaniasis is a protozoan disease whose diverse clinical manifestations are dependent both on the infecting species of Leishmania and the immune response of the host. Transmission of the disease occurs by the bite of a sand fly infected with Leishmania parasites. Infection may be restricted to the skin in cutaneous leishmaniasis, limited to the mucous membranes in mucosal leishmaniasis, or spread internally in visceral leishmaniasis or kala azar. The overall prevalence of leishmaniasis is 12 million cases worldwide, and the global yearly incidence of all clinical forms approaches 2 million new cases (World Health Organization WHO/ LEISH/200.42, Leishmania/HIV Co-Infection in Southwestern Europe 1990-98: Retrospective Analysis of 965 Cases, 2000). In the last two decades, leishmaniasis, especially visceral leishmaniasis, has been recognized as an opportunistic disease in the immunocompromised, particularly in patients infected with human immunodeficiency virus.

Expression of fos and jun proto-oncogenes in benign versus malignant human uterine tissue.

OBJECTIVE: The objective of this study was to evaluate expression of fos and jun proto-oncogenes in benign human uterine tissue compared with malignant uterine tissue. METHODS: Forty-two endometrial tissue specimens were obtained at the time of hysterectomy. Tissue samples from different phases of the menstrual cycle and from postmenopausal patients were stained using immunohistochemical methods to detect Fos and Jun proteins, estrogen and progesterone receptor status, and Ki67 (detects a nuclear antigen associated with proliferating cells). Tissue was examined microscopically for nuclear staining in endometrial epithelium and stroma. The endometrium was based on the patient's last menstrual period, pathologic dating, and proliferative versus nonproliferative status as determined by Ki67. Benign and malignant specimens were subjected to Northern blot analysis to evaluate levels of expression of c-fos, c-jun, and jun-B mRNA. The pattern of c-fos mRNA expression in malignant samples was further evaluated using in situ hybridization. RESULTS: In proliferative, secretory, postmenopausal, and progesterone-influenced, uterine specimens immunohistochemically stained and examined, the endometrial and stromal nuclei stained for both Fos and Jun in varying intensities. However, no pattern was found in the variation of intensity according to the phase of the endometrium. Similarly, in malignant and benign endometrial tissue examined by Northern blot and in situ hybridization analyses, expression of proto-oncogene mRNAs was readily detectable, but no statistical correlation between type of tissue examined, grade of adenocarcinoma, and stage of endometrial cancer was found in this study. CONCLUSIONS: In rodent models, control of uterine cell proliferation is related to change in expression of fos and jun proto-oncogenes. Our results indicate that hormonal control is likely to be different in human endometrium and probably involves genes other than the proto-oncogenes under study. Expression of Fos and Jun do not correlate with endometrial cancer stage and grade.

Effects of ethanol on lateral and rotational mobility of plasma membrane vesicles isolated from cultured mouse myeloma cell line Sp2/0-Ag14.

Intramolecular excimerization of Py-3-Py and fluorescence polarization of DPH were used to evaluate effects of ethanol on the rate and range of the lateral mobility and the range of the rotational mobility of bulk bilayer structures of the Sp2/0-PMV. In a concentration-dependent manner, ethanol increased the rate and range of the lateral mobility and the range of the rotational mobility of bulk bilayer structures of Sp2/0-PMV. Selective quenching of DPH by trinitrophenyl groups was utilized to examine the range of transbilayer asymmetric rotational mobility of the Sp2/0-PVM. The anisotropy (r), limiting anisotropy (r(infinity)) and order parameter (S) of DPH in the inner monolayer were 0.022, 0.029 and 0.063, respectively, greater than calculated for the outer monolayer of the Sp2/0-PMV. Selective quenching of DPH by trinitrophenyl groups was also used to examine the transbilayer asymmetric effects of ethanol on the range of the rotational mobility of the Sp2/0-PMV. Ethanol had a greater increasing effect on the range of the rotational mobility of the outer monolayer as compared to the inner monolayer of the Sp2/0-PMV. It has been proven that ethanol exhibits a selective rather than nonselective fluidizing effect within the transbilayer domains of the Sp2/0-PMV.

Inhibin suppresses human chorionic gonadotropin secretion in term, but not first trimester, placenta.

Inhibin is produced by the placenta, with serum concentrations rising throughout pregnancy. In contrast, hCG serum concentrations peak in the first trimester and are 80-90% lower at term. This study was designed to determine the effect of inhibin on hCG secretion both early and late in gestation. Villus tissue from 3 term and 3 first trimester (8-10 week) placentas was maintained in an in vitro explant culture model for 5 days. Tissue from each placenta was incubated with control medium in 24 replicate wells for the first 72 h. During the final 48 h, 12 wells received control medium, and 12 wells received medium containing 1% rabbit antiserum raised against the alpha-subunit (residues 1-32) of the human inhibin peptide. The antiserum demonstrated biological activity by increasing serum FSH concentrations in an immature female rat bioassay. The relative increase in hCG secretion at the conclusion of days 4 and 5 in control and antiserum-treated groups for each first trimester and term placenta were compared to pretreatment hCG concentrations on day 3. The relative increases in hCG secretion of first trimester control groups on day 4 (mean +/- SD, 34 +/- 11%) and day 5 (63 +/- 23%) were compared to those in antiserum-treated groups on day 4 (39 +/- 13%) and day 5 (54 +/- 5%) and showed no significant difference between groups on either day. The same comparison in term cultures showed the relative increases in hCG secretion of control groups on day 4 (31 +/- 10%) and day 5 (64 +/- 50%) to be significantly lower than those in antiserum-treated groups on day 4 (100 +/- 41%) and day 5 (150 +/- 108%; P less than 0.001). These findings suggest that inhibin suppresses hCG secretion in term, but not first trimester, placentas.