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      N-acetyl-cysteine mediates protection against Mycobacterium avium through induction of human β-defensin-2 in a mouse lung infection model

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      Microbes and Infection
      Elsevier BV

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
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            Epidemiology of Pulmonary Nontuberculous Mycobacterial Disease, Japan1

            To the Editor: Incidence of pulmonary nontuberculous mycobacterial disease (PNTMD) is reportedly increasing globally ( 1 , 2 ). Although such an increase is expected in Japan ( 3 , 4 ), the epidemiologic situation is unclear. The most recent survey, which used the 1997 American Thoracic Society diagnostic criteria, reported that the incidence rate for PNTMD in 2007 was 5.7 cases per 100,000 person-years ( 5 ). To update the data, we performed a nationwide hospital-based survey in Japan. After a preliminary survey of 20 hospitals, we developed and disseminated questionnaires to all 884 hospitals in Japan that were certified by the Japanese Respiratory Society. The surveys asked about the number of newly diagnosed cases, from January through March 2014, of PNTMD, pulmonary Mycobacterium avium disease, M. intracellulare disease, or M. avium complex (MAC; the combination of the first 2 species listed); pulmonary M. kansasii disease; pulmonary M. abscessus disease; and tuberculosis (TB) for inpatients and outpatients. Hospital respondents returned the completed questionnaires by mail, fax, or Internet. To avoid potential reporting bias and misclassification, we counted only cases that met the 2007 American Thoracic Society/Infectious Diseases Society of America statements ( 6 ) and excluded cases diagnosed at other hospitals. Because the source population can be ascertained by using the epidemiologic data for TB as a reportable disease, to estimate the incidence rate of PNTMD, we used the ratio of TB to PNTMD cases. The PNTMD incidence rate was calculated as the national incidence rate of TB multiplied by the ratio of new PNTMD to TB cases reported by the responding hospitals (online Technical Appendix Figure 1, http://wwwnc.cdc.gov/EID/article/22/6/15-1086-Techapp1.pdf). To clarify the chronologic changes in incidence, we followed the same method for comparing TB and PNTMD used in a prior epidemiologic study in Japan ( 5 ). We established methods for maximizing survey response rates and facilitating ease of completion by offering extensive support to survey recipients (Technical Appendix Table 1). We achieved a high response rate of 62.3% (551 hospitals), and in all regions the response rate exceeded 50% (Technical Appendix Table 2). The numbers of newly diagnosed cases were 2,327 for TB and 2,652 for PNTMD. Because the incidence rate for TB was reported to be 12.9 cases per 100,000 person-years, that of PNTMD was estimated to be 14.7 cases per 100,000 person-years, which is ≈2.6 times the incidence rate reported in 2007 (Figure). By using the same method, we found the incidence of pulmonary MAC, M. kansasii, and M. abscessus disease to be 13.1, 0.6, and 0.5 cases per 100,000 person-years, respectively (Technical Appendix Table 2). The ratio of pulmonary M. avium disease to MAC was higher in the northern and eastern parts of Japan, whereas the ratio of pulmonary M. intracellulare disease to MAC was higher in the southern and western parts of Japan (Technical Appendix Figure 1). Figure Incidence (no. cases/100,000 person-years) of pulmonary nontuberculous mycobacterial (NTM) disease, culture-positive tuberculosis (TB), and smear-positive TB in Japan during 1980–2014. The nationwide survey revealed that the incidence rate of pulmonary NTM disease exceeds that of TB. The epidemiologic survey before 1988 was conducted annually by the same research group; subsequently, another group performed the epidemiologic survey only in 2001 and 2007. From this survey, we observed that the incidence rate of PNTMD may exceed that of TB and that incidence rates of PNTMD in Japan may be among the highest worldwide (Figure). This finding implies that the prevalence of PNTMD as a chronic infection is estimated to be much higher than that of TB. We assume that the high rates of PNTMD in Japan are consistent with data suggesting that Asians are particularly susceptible to PNTMD ( 1 , 7 , 8 ). Other factors contributing to the increase might be the simplified diagnosis according to the 2007 American Thoracic Society/Infectious Diseases Society of America statements, increased awareness by medical staff, population aging, and increased frequency of medical checkups with computed tomography of the chest. Another finding was the characteristic gradient clustering of the ratios of M. avium and M. intracellulare (Technical Appendix Figure 2). This finding supports the widely accepted belief that environmental factors strongly affect the epidemiology of PNTMD; therefore, the role of factors such as soil, humidity, temperature, and saturated vapor pressure should be seriously considered ( 9 ). We also found dramatic increases in incidence of pulmonary M. abscessus disease and pulmonary MAC disease, whereas incidence of pulmonary M. kansasii disease was stable. Although we did not distinguish M. massiliense from M. abscessus, the incidence rate for pulmonary M. abscessus disease increased from 0.1 cases in 2001 to 0.5 cases per 100,000 person-years in 2014. This epidemiologic tendency should be monitored ( 10 ). This study has several limitations. First, differing characteristics between the responding and nonresponding hospitals could cause bias. Second, we did not collect data outside of hospitals. Third, incomplete reporting could undermine the accuracy of our estimates (Technical Appendix Tables 3, 4). Therefore, the epidemiologic data should be verified by using other approaches (Technical Appendix Table 1). The dramatic increase in incidence rates for PNTMD warrants its recognition as a major public health concern. Because the prevalence rates of this currently incurable lifelong chronic disease are estimated to be high, the effect on the community could be enormous. Further investigations are needed. Technical Appendix Incidence rates for mycobacterial infections in Japan during 1980–2014; response rate, results, characteristics, and limitations of survey of newly diagnosed pulmonary nontuberculous mycobacterial disease and mycobacterial disease, January–March 2014, Japan; and comparison of hospitals that did and did not respond to the survey.
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              Mammalian defensins in the antimicrobial immune response.

              Defensins are peptidic components of the innate immune system of plants and animals. In mammals, defensins have evolved to have a central function in the host defense properties of granulocytic leukocytes, mucosal surfaces, skin and other epithelia. This review focuses on the biological functions of three structural subgroups of mammalian defensins and the evidence for their involvement as effectors of antimicrobial innate immunity.
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                Author and article information

                Contributors
                (View ORCID Profile)
                Journal
                Microbes and Infection
                Microbes and Infection
                Elsevier BV
                12864579
                November 2020
                November 2020
                : 22
                : 10
                : 567-575
                Article
                10.1016/j.micinf.2020.08.003
                32882411
                803d3eec-86ec-4fe9-b5f5-111ccfea0924
                © 2020

                https://www.elsevier.com/tdm/userlicense/1.0/

                http://creativecommons.org/licenses/by/4.0/

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